CZ372096A3 - Compounds and methods of treating cardiovascular, respiration and immunity complaints - Google Patents

Abstract

Tetrahydrofurans, tetrahydrothiophenes, pyrrolidines and cyclopentanes are disclosed that reduce the chemotaxis and respiratory burst leading to the formation of damaging oxygen radicals of polymorphonuclear leukocytes during an inflammatory or immune response. The compounds exhibit this biological activity by acting as PAF receptor antagonists, by inhibiting the enzyme 5-lipoxygenase, or by exhibiting dual activity, i.e., by acting as both a PAF receptor antagonist and inhibitor of 5-lipoxygenase. It has been determined that 5-lipoxygenase activity, oral availability, and stability in vivo (for example, glucuronidation rate) can vary significantly among the optical isomers of the disclosed compounds.

Description

COMPOUNDS AND METHODS FOR THE TREATMENT OF CARDIOVASCULAR, BREATHING, AND IMMUNE DISEASES i X 'AREA OF THE INVENTION v; \ - i ,. ¹ i ”. * ..: »σ.

The present invention is in the field of 2,5-disubstituted tetrahydrothiophenes, pyrrolidines and 1,3-disubstituted cyclopentanes. These compounds exhibit biological activity by inhibiting the 5-lipoxygenase enzyme, acting as PAF receptor antagonists or dual activity by acting as PAF receptor antagonists and 5-lipoxygenase inhibitors. ,

BACKGROUND OF THE PROBLEM

Leukotrienes are potential local mediators playing a major role in respiratory and allergic responses including arthrosis, asthma, psoriasis and thrombotic diseases. Leukotrienes are straight-chainedosanoic chains formed as a product of the oxidation of acidicosanoic acid by lipogenases. Icosanoic acid is oxidized by 5-lipoxygenase to hydroperoxido 5-hydroperoxy-icosatetraenoic acid (5-HPETE), which is converted to leukotriene A4, which in turn can be converted to leukotriene B4, C4 or D4. The slow reacting substance of anaphylaxis is now known as a mixture of C4, D4, and E4 leukotrienes, each of which is a potent limiter of bronchitis. There is research to develop specific receptor antagonists or inhibitors of leukotriene biosynthesis to prevent or minimize pathogenic respiratory responses mediated by these compounds. ,. European Patent Nos. 90117171.0 and 901170171.0 have disclosed that indole, benzofuran and benzothiophene are lipoxygenase inhibiting compounds.

More recently, thiophene derivative L has been shown to be a hydrophthalic anhydride _; 652,731, trans-2,5-bis- (3,4,5-trimethoxyphenyl) tetrahydrothiophene (L-653,150) is strong. a PAF antagonist; and a mild 5-lipoxygenase inhibitor. Certain 2,5diaryl-tetradydrothiophenes have been reported to be PAF antagonists and inhibitors of leukotriene synthesis.

(Biftu, et al., Abstract. 6th International Conference on Prostaglandins and Related Compounds, June 3-6, 1986, Florence, Italy; U.S. Patent No. 4,757,084 to Biftu); WO 92/15294; WO 94/01430; WO 94/04537 and WO 94/06790.

WO 92/13848 discloses a class of racemic hydroxamic acids and N-hydroxyureas which inhibit lipoxygenase having the structure:

wherein R ¹ is hydrogen, alkyl, alkenyl, amine or substituted amine, R ⁴ is hydrogen, a pharmaceutically acceptable cation, aroyl or alkoyl, A is alkylene or alkenylene, X is oxygen or sulfur, each Y is hydrogen, halogen, cyano, hydroxy, alkyl, alkoxy, alkylthio, alkenyl, alkoxyalkyl, cycloalkyl, aryl, aryloxy, arylalkyl, arylalkenyl, arylalkoxy or substituted aryl, Z is oxygen or sulfur, m is 0 or 1, n is 1-5 and p is 2-6.

Assuming a significant number of pathological immune and respiratory responses that are mediated by 5-lipoxygenase, there remains a need to identify novel compounds and compositions that inhibit this enzyme.

Platelet activating factor (PAF, 1-O-alkyl-2-acetyl-1'-glycerol-3-phosphorylcholine) is a potent respiratory phospholipid mediator of a wide range of biological activities. PAF was originally identified as a water-soluble compound released by immunoglobulin E (IgE) - a senitized rabbit basophil. It is now known that PAF is also generated and released by monocytes, macrophages, polymorphonuclear leukocytes (PMNs), icosanophils, neutrophils, natural lymphocyte destroyers, platelets, and endothelial cells, as well as kidney and cardiac tissues under appropriate immunological and non-immunological conditions. (Hwag, “Specific receptors for platelet activating factor, receptor heterogeneity and trans-

duction ”, Journal of Lipid Mediators 2,123 (1990)). PAF causes the aggregation and gegranulaci platelets at low concentration levels in tissues efficiency (active at 10 ^-12 to 10 @ ^-9 M, picomoles) and short plasma life time (2-4 minutes) of PAF are similar to those that occur naturally other fat mediators such as thromboxane A ₂ , prostaglandins and leukotrienes.

PAF mediates biological responses by binding to specific receptors found in a wide variety of cells and tissues. Studies on the effect of structure on the activity of PAF and analogs suggest that the ability of PAF and its analogs to bind these receptors is structurally specific and stereospecific. (Shen, et al., "Chemical and Biological Properties of PAF Antagonists and Biosynthetic Inhibitors," Factor Activating Plates and Related Fat Mediators, F. Snyder, Ed. Plenum Press, New York, NY 153 (1987)).

Although PAF mediates substantial biological responses, it also appears to play a role in pathological immune and respiratory responses. Many published studies have provided evidence that PAF is involved in human diseases including arthrosis, acute inflammation, asthma, endotoxic shock, pain, psoriasis, eye inflammation, ischemic heart disease, intestinal ulceration, myocardial infarction, rectal inflammation and acute respiratory syndromes. . Animal experiments also show that PAF is produced or increases in certain pathological conditions.

The presence of PAF in pathological respiratory and immune conditions stimulates significant research to identify PAF receptor antagonists. In 1983, it was reported that the phospholipid analog, described as CV-3988 (rac-3- (N-noctadecylcarbamoyloxy-t-methoxypropyl-2-thiazolioethyl phosphate), has properties as a PAF receptor antagonist (Terashita, et al. , Life Sciences 32, 1975 (1983).

In other early works in this field, Shen, et al. (In Proc. Natl. Acad, Sci (USA) 82, 672 (1985)) reported that cadsurenone, a neolignan derivative isolated from Piper fútokadsura Sieb et Zucc (Chinese plant) ), is a potent, specific, and competing inhibitor of PAF activity at the receptor level.

Hwang, et al., Reported in 1985 that trans-2,5-bis- (3,4,5-trimethoxyphenyl) tetrahydrofuran (L-652,731) inhibited the binding of tritiated PAF to the PAF receptor sites. (Hwang, et.al., " Ya-77.y-2,5-bis (3,4,5-trimethoxyphenyltetrahydrofuran "), Journal of

I · · * * · * * «« «« «« «« «« «« «« «« I · ···········

In Biological Chemistrv 260,15639 (1985), L-652,731 showed that it is orally active and that it inhibits rat skin permeability induced by PAF at a dose of 30 mg / kg body weight. This compound has been shown to have no effect on the 5-lipoxygenase enzyme. Hwang, et.al. have also shown that fAny-L-652,731 (in which the aryl groups at positions 2 and 5 on opposite sides of the plane of the tetrahydrofuran ring) is approximately 100 times stronger than cis-L-652,731 (in which substituents 2 and 5 are on the same side of the plane of tetrahydrofuran) circle).

In 1988, Hwang, et al. Reported that L-659,989 (trans-2- (3-methoxy-4-propoxyphenyl-5-methylsulfonyl) -5- (3,4,5-trimethoxyphenyl) tetrahydrofuran is orally

J active, potent, PAF receptor antagonist with an inhibition constant 10 times greater than trans. L-652.73. (Hwang, et al., J. Pharmacol. Ther. 246,534 (1988)).

U.S. Pat. patents 4,996,203; Nos. 5,001,123 and 4,539,332 to Bifla, et al. And European Patents Nos. 8902593.3, 90306235.4 and 90306234.7 disclose that specific classes of 2,5-diaryltetrahydrofurans are PAF receptor antagonists.

Bowless et al., Svnlett, 1993, pp 111 disclose and limited the number of substituted tetrahydrofuran which may exhibit receptor antagonism. against PAF.

Danyoshi et al., Chem.Pharm.Bull, 1989 p. 1969 published 2-substituted Nalkoxycarbonylpyrrolidines which inhibit PAF-induced rabbit platelet aggregation.

It is therefore an object of the present invention to provide compounds that reduce chemotaxis and respiratory attacks that result in the formation of harmful oxygen radicals during respiratory or immune responses.

Another object of the present invention is to provide pharmaceutical compounds for the treatment of pathological immune or respiratory problems mediated by lipoxygenase products.

Another object of the present invention is to provide a method for treating pathological immune or respiratory problems caused by 5-lipoxygenase products.

Yet another object of the present invention is to provide pharmaceutical compounds for the treatment of pathological immune or respiratory problems caused by PAF.

SUMMARY OF THE INVENTION • V Cl f CC fc V «f f '' '' '' '' '* * p p p p p 4 f f 4 4 4 V ': forr c-1 cv <c *' at $ 4

Compounds of formula I are provided

(I) wherein;

Άτ is an aryl or aetheroaryl group which is cast optionally substituted, preferably by halogen (including but not limited to fluorine), lower alkoxy (including methoxy), lower aryloxy (including phenoxy), W, cyano or R ³ ; 'L'.

m is 0 or 1;

q is 0 or U; '' · ' ¹ ' - '*' in J.

¹ tn is 0-6; · «. '·''> · · ⁴ ' V 'iv. W is independently -AN (OM) C (O) N (R ³⁾ R ^{4, 1} -N (OM) C (O) N (R ³⁾ R ⁴ .- AN (R ³⁾ C (O) N (OM) R ⁴ , -N (R ³ ) C (O) N (OM) R ⁴ , -AN (OM) C (O) R ⁴ , -N (OM) C (O) R ⁴ -, AC ( O) N (OM) R ⁴ , -C (O) NHA or -AB;

A is lower alkyl, lower alkenyl, lower alkynyl, alkylaryl or arylalkyl, in which one or more carbons may be optionally replaced by O, N, or sulfur (supplemented with oxygen or hydrogen, as necessary), but Y-Α-, -A-, or -AW- should not contain two adjacent heteroatoms (such as -00, -SS, -Ο-S-, etc.) (In one example, the lower alkyl is a bridged alkyl group such as eg - (CH 2) _n C (lower alkyl) H-, wherein n is 1-5, and specifically - (CH 2) C (CH 3) H - or lower * alkynyl of the formula -G 5 C-CH (lower alkyl) -, including - C≡C-CH _(CH3) -);

The group Β is selected from the group consisting of pyridylimidazole and benzimidazole, both of which are substituted with R ³ , and wherein the pyrridylimidazole or benzimidazole are preferably linked via a nitrogen atom;

M is hydrogen, a pharmaceutically acceptable cation, or a metabolically cleavable group;

X is O, S, S (O), S (O) ₂ , NR ³ , or CHR ⁵ ;

Y is O, S, S (O), S (O) ₂ , NR ³ , or CHR ⁵ ;

Z is O, S, S (O), S (O) ₂ , NR ³ ;

R and R j are independently hydrogen, lower alkyl including methyl LU Dimethyl cyclopropyl, ethyl, isopropyl, butyl, pentyl, hexyl, and C ₃ .g cycloalkyl, e.g. cyclopentyl; haloalkyl such as trifluoromethyl; halogen, e.g. fluorine; and COOH;

R ³ and R ⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl, arylalkyl, alkylaryl, C 1-6 alkoxy- C 1-6 alkyl, C 1-6 alkylthio-C 1-6 alkyl, heteroaryl, or hetero arylalkyl-;

R ⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, arylalkyl, alkylaryl, AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , -AN (OM) C (O) R ^4, -AC (O) N (OM) R ^4, S (O) x R ^3, -AS (O) 'CH 2 C (O) R ^3, -AS (O) _n CH ₂ CH (OH) R ³ or -AC (O) NHR ³ where x is 0 2.

The group Ar is in one instance selected from the group consisting of phenyl, trimethoxyphenyl, dimethoxyphenyl, fluorophenyl (especially 4-fluorophenyl), difluorophenyl, pyrrolidine, dimethoxypyrridyl, quinolinyl, furyl, imidazolyl.

In one instance, -A-B is w

□. · »*. ·· “'· | - - I JI *. X · —M.

and Ar is an optionally substituted aryl or heteroaryl group, as described in more detail in section I.A. below.

13: i? Ί 0 * 3

OC. d C C <*;

13 · fc O Č C ^! d or> ccc O 'cc <c

CO ΓΤ 0 ** <: · π <!

Ir ft

IS 'fl 0 <?:

(i) <»'*« <: f r »';

m v «ί.

* 4: iJ c f ♦?

Non-limiting examples of preferred compounds are:

wherein R ¹⁰ is halogen, -CN, hydrogen, lower alkyl, lower alkenyl, lower alkynyl, lower alkoxy or lower aryloxy. + '_ ^v · _;

K ^r These compounds in general reduce the chemotaxis and respiratory burst leading to the formation of harmful radicals of polymorphonuclear leukocytes during an inflammatory or> immune responses. These compounds generally exhibit biological activity by inhibiting the 5-lipoxygenase enzyme, playing the role of PAF receptor antagonists or by exhibiting so-called dual activity, i.e. acting both as a PAF receptor antagonist and a 5-lipoxygenase inhibitor.

Another embodiment of the invention is a pharmaceutical composition comprising an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or derivative thereof, in combination with a pharmaceutically acceptable carrier, for any of the disorders described above.

In a preferred embodiment, the compounds are used to treat disorders. mediated by 5-lipoxygenase, by administering an effective amount of one or more of the previously described compounds or pharmaceutically acceptable salts or derivatives, which is any pharmaceutically acceptable carrier.

Surprisingly, it has been determined that the activity of a compound, eg, 5-lipoxygenase activity, oral availability and in vivo stability (eg, glucuronide ratio), can vary significantly when switching from one optical isomer to another. Therefore, in one embodiment of the invention, the compound is administered in enantiomerically enriched form.

Examples of immune, allergic and cardiovascular disorders include general inflammation, cardiovascular disorders including hypertension, bone and muscle problems, bone arthritis, gout, asthma, pulmonary edema, adult respiratory syndromes, pain, platelet aggregation, shocks, rheumatoid arthritis, rheumatoid arthritis, juvenile arthritis arthritis, psoriasis, autoimmune uveitis, allergic encephalomyelitis, systemic lupus erythematosis, acute necrotizing, haemorrhagic encephalopathy, idiopathic thrombocytopenia, polychondritis, chronic active hepatide, idiopathic maladsorption syndrome, cirrhosis, Crohn's disease, Crohn's disease, Crohn's disease allergic asthma, inappropriate allergic responses to? environmental stimuli such as ivy poisoning, pollen, insect stings, certain foods, including atopic dermatitis and contact dermatitis.

The compounds disclosed herein may also be used as research tools to study biological pathways including leukotrienes or PAF, if appropriate.

The following examples are non-limiting examples of compounds that fall within the scope of compounds having Formula I. They are exemplary only and are not intended to exhaust the entire scope of the invention.

trans-2- (3,4,5-trimethoxyphenoxymethyl) -5- [4-N '-methyl-N' hydroxyuridyl) butyl] tetrahydrofuran;

trans-2- (3,4,5-trimethoxyphenoxymethyl) -5- (4-N '-methyl-N' -hydroxyireidyl) -butinyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N' -hydroxyureidyl) butyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N'-methyl-N'-hydroxyureidyl) -1-butynyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-butyl-N' - (hydroxyureidyl) butyl] tetrahydrofuran;

_f trans-2- (4-fIuorofenoxymethyl) -5- [4-N'-butyl-N'-hydroxyureidyl) l-butynyl] tetrahydrofuran;

trans-2- (3,4,5-trimethoxyphenoxymethyl) -5- [4-N '-methyl-N-hydroxyureidyl) butyl] tetrahydrofuran;

Φ Φ · trans------------------------- 3,4,5-trimethoxyphenoxymethyl) -5- [4-N'-methyl-N-hydroxyureidyl) butynyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N-hydroxyuridyl) butyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N-hydroxyureidyl) -1-butynyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-butyl-N-hydroxyureidyl) butyl] tetrahydrofuran;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-butyl-N-hydroxyureidyl) -1-butynyl] tetrahydrofuran;

trans-2- _(3,4} 5-trimethoxyfenoxymethyl) -5- [4-N'-Methyl-N'hydroxyureidyl) butyl] tetrahydrothiophene;

trans-2- (3,4,5-trimethoxyphenoxymethyl) -5- [4-N '-methyl-N'-hydroxyureidyl) 1 butynylphetrahydrothiophene;

trans -2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N' -hydroxyureid 1) butyl] tetrahydrothiophene;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N' -hydroxyureidyl) -1-butyl 1] tetrahydrothiophene;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-N' hydroxyureidyl) butyl] tetrahydrothiophene;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-butyl-N' -hydroxyureidyl) 1 -butyl 1) tetrahydrothiophene;

trans-2- (3,4,5-trimethoxyphenoxymethyl) -5- [4-N '-methyl-N-hydroxyureidyl) butyl] tetrahydrothiophene;

trans-2- (3,4,5-trimethoxyphenoxymethyl) -5- [4-N '-methyl-N-hydroxyurea 1) 1 butynyl] tetrahydrothiophene;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N-hydroxyureidyl) butyl].

tetrahydrothiophene;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-methyl-N-hydroxyureidyl) -1-butynyl] tetrahydrofuran;

• «« «

Trans-2- (4-fluorophenoxymethyl) -5- [4-N'-butyl-N-] trans-2- (4-fluorophenoxymethyl) -5- [4-N'-butyl-N-] hydroxyureidyl) butyl] tetrahydrothiophene;

trans-2- (4-fluorophenoxymethyl) -5- [4-N '-butyl-N-hydroxyureidyl) -1-butynyl] tetrahydrothiophene;

2- (3,4,5-trimethoxyphenyl) -5- [3- (N'-methyl-N'-hydroxyureidyl) propoxy] tetrahydrofuran;

2- (4-fluorophenyl) -5- [3- (N'-methyl-N'-hydroxyureidyl) propoxy] tetrahydrofuran;

2- (3,4,5-trimethoxyphenyl) -5- [3- (N'-n-butyl-N'-hydroxyureidyl) propoxy] tetrahydrofuran;

2- (4-fluorophenyl) -5- [3- (N'-n-butyl-N'-hydroxyureidyl) propoxy] tetrahydrofuran;

2- (3 &apos;, 4 &apos; -methoxyphenyl) -5- [3- (N-butyl-N-hydroxyureidyl)] propoxytetrahydrofuran;

- (3 4, 4 im-dimethoxyphenyl) -5- [3-N-methyl-N-hydroxyroxide 1) propoxytetrahydrofuran;

2- (2,4,5-trimethoxyphenyl) -5- (3-hydroxyureidylpropoxy) tetrahydrofuran; 2- (4-fluorophenyl) -5- (3-hydroxyureidylpropoxy) tetrahydrofuran; 2- (4-fluorophenyl) -5- [3- (’-methyl-1'-hydroxyhydroxyureidyl) propoxy] tetrahydrothiophene; and 2- (4-fluorophenyl) -5- [3hydroxyureidylpropoxy) tetrahydrothiophene.

Other non-limiting examples of other compounds of Formula I are given in the tables

1, 2 and 3 and in figures 1a and 1b.

·>

• «4 «4 • • » 44 • 4 4 4 · ·· 4 · 4- 4 44 4 4 · 4 4 4 44 • • 4 • 4 * 4 4 4 »44 44 444 4 « 4 4

TABLE ί

Aí * X w XJ 0 CH 3 CH, CH, NHC (O) H (OH) Cl 2 C SAME SAME with SAME SAME NH SAME SAME 0 CH3CH3CH, N (OH) (CCO) Mt SAME C SAME SAME with SAME SAME NH SAME SAME 0 CH ^CHjCHjMfOHJCtOJNHCh SAME C SAME SAME 0 r v. Mí jwMAYqu. SAME AS ABOVE SAME AS ABOVE

^X 0 '

SAME AS ABOVE SAME AS ABOVE ₀ AND _n ^ SAME AS ABOVE SAME AS ABOVE

SAME AS ABOVE SAME AS ABOVE

C refers to CHR ⁵ . Y is O, CHR ⁵ , S or NH.

 «4 * v • • • · · < • ·  · • • · «# · * · • • and • in »« • • W J · • • ·· • * ··· « fc • ·

TABLE 2

Ar

Ar

W

F

same as above

same as above

- 9 *

QH same as above

F

»·· • »» # • » • • · ♦ ·· • ·  · • • ··· • · • « • ··· • • * • • • • • • · * « ·· ··· • · ··

TABLE ΙΪΙ · ίη. ·· .ν

XYU-rt

OH

AND

Ν NH,

OH

CH, _O

N NH,

S _> _CH3

- ¢ - _Q 'nh,

OH ²

-.-CH,

1 ^ -A 9

OH ^NH >

-O- ™ .- jy ^and ^ ° rv ° '- ^' mr ^w

-Ό * and 'O' CCm R

Ι ^ ΆΛ

OH

OH

OH l

NH,

NH,

OH

NH, yy, phyphs

°'ΎαΓ o '* σκτ .w ° A \ < ^w

R R jy ° - .O fyn oOCUT

• a · • · «A · * · • a • and and and a • · a · * · • • a and a and a • « • and • and a • a a · • a • a · • · and a

BRIEF DESCRIPTION OF THE FIGURE

Figures 1a and 1b illustrate chemical structures with indicated stereochemistry of selected active compounds.

Figure 2 illustrates the degree of glucuronidation of racemic compound 202 as well as its enantiomers, as well as compounds 216,217,234 and 236.

Figure 3 illustrates the degree of glucuronidation of the following enantiomers shown.

Figure 4 is an illustration of the synthesis process of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5 '(4-N-hydroxyureidyl-1-butynyl) tetrahydrofuran (compound 401) and 2S, 5R-trans-2- (4-fluorophenoxymethyl) -5- (4-N-hydroxyureidylbutyl) tetrahydrofuran (Compound 402).

Figure 5 is a graph of the glucuronidation ratio of compounds 401,403,404 and 406 (as shown in Table 4) measured in percent metabolite versus time (in hours).

A a · a a a a a a a a a a a a a a

DETAILED DESCRIPTION OF THE INVENTION

I. Description and synthesis of compounds

Compounds A

As used herein, the term "enantiomerically enriched" refers to a compound that exhibits a form comprising at least 95%, preferably 97%, 98%, 99.% or 100%, of a single enantiomer of the compound.

The term alkyl, as used herein, and unless otherwise specified, refers to a saturated, straight, branched, or cyclic C 1 -C 10 hydrocarbon, specifically including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, 3-methylpentyl, 2,2-dimethylbutyl and 2,3-dimethylbutyl. The alkyl group may be optionally substituted with a suitable group including, but not limited to, R ³ , or one or more selected from halogen, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acids, sulfate, phosphoric acid, or phosphate, either protected or unprotected, as is known to those skilled in the art, as taught in Greene, et al., "Protecting Groups in Organic Chemistry," John Wiley and Sons, Second Edition, 1991.

The term halogen as used herein refers to chlorine, fluorine, bromine and iodine.

The term lower alkyl as referred to herein and unless otherwise specified refers to a C 1 -C 6 saturated straight, branched, or cyclic (in the case of C 5-6) hydrocarbon, specifically including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, 3-methylpentyl, 2,2-dimethylbutyl and 2,3-dimethylbutyl, optionally substituted as described above for an alkyl group.

The term alkenyl, as referred to herein, and unless otherwise specified, refers to a straight, branched, or cyclic (in the case of C 1-6) C ₂ to C 10 hydrocarbon with at least one single double bond, optionally substituted as used herein it is referenced above.

· * «· · · · · · · * * * * * * *

The term lower alkenyl as referred to herein and unless otherwise indicated refers to an alkenyl group of C ₂ -C 6, specifically includes vinyl and allyl.

The term lower alkylamino refers to an amino group having one or two alkyl substituents.

The term alkynyl, as referred to herein, and unless otherwise indicated, refers to a straight or branched C ₂ to C 10 hydrocarbon having at least one triple bond, optionally substituted as described above. The term lower alkynyl, as referred to herein, and unless otherwise specified, refers to a C ₂ to C alkynyl group, specifically including acethylenyl, propynyl, and -C? C-CH (CH3) ~.

The term aryl as used herein and unless otherwise indicated refers to phenyl, biphenyl or naphthyl, but phenyl is preferred. The aryl group may be optionally substituted with a suitable group including, but not limited to, one or more moieties selected from the group consisting of halogen, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acids, sulfate, phosphoric acid , phosphate or phosphite, protected or unprotected, as known to those skilled in the art, such as taught in Greene, et al., "Protecting Groups in Organic Synthesis", John Wiley and Sons, Second Edition, 1991 but preferably from halogen (including, but not only from fluoro), lower alkyloxy (accounting methoxy), lower aryloxy (accounting phenoxy), W, cyano or R ^3rd

The term haloalkyl, haloalkenyl, or haloalkynyl refers to an alkyl, alkenyl or alkynyl group in which at least one hydrogen of the group is replaced by a halogen atom.

The term heteroaryl, heterocycle or heteroaromatic, as referred to herein, refers to an aromatic moiety that contains at least one sulfur, oxygen, or nitrogen in the aromatic ring, which may be optionally substituted as described for aryl groups. Examples include, but are not limited to, pyryl, furyl, pyridyl, 1,2,4-thiadiazolyl, pyrimidyl, thienyl, isothiazolyl, imidazolyl, tetrazolyl, pyrazole, pyrimidyl, quinolyl, isoquinolyl, benzothienyl, isobenzofuryl, pyrazolyl, indolyl, purinyl, carbazolyl, benzimidazolyl and isoxazolyl.

The term aralkyl refers to an aryl group with an alkyl substituent.

** * ** ** · ** ** ** ** «*« * * * * er er er er er er er er er er * »

The term alkaryl refers to an alkyl group having an aryl substituent.

The terms organic and inorganic anion refer to an organic and inorganic group that carries a negative charge and may represent a negative portion of the salt.

The term "pharmaceutically acceptable cation" refers to an organic or inorganic grouping of atoms that bears a positive charge and which can be administered in conjunction with a pharmaceutical agent, such as a counterion in a salt.

Pharmaceutically acceptable cations are known to those skilled in the art and include, but are not limited to, sodium, potassium, and quaternary amines.

The term "metabolically cleavable group" refers to a moiety that can be cleaved in vivo from a molecule to which it is bound and includes, but is not limited to, an organic and inorganic anion, a pharmaceutically acceptable cation, an acyl (e.g., (alkyl) C (O) , including acetyl, propionyl and butyryl), alkyl, phosphate, sulfate and sulfonate.

The term PAF receptor antagonist refers to a compound that binds to a PAF receptor with a binding constant of 30μΜ or less.

The term 5-lipoxygenase inhibitor refers to a compound that inhibits an enzyme at 30 μΜ or less in a disrupted cellular system.

The term pharmaceutically active derivative refers to any compound which, when administered to a recipient, is capable of providing directly or indirectly disclosed compounds,

The 2,5-disubstituted tetrahydrothiophenes, tetrahydrofurans and pyrrolidines as well as the 1,3-disubstituted cyclopentanes described herein exhibit PAF receptor antagonist activity, inhibit the 5-lipoxygenase enzyme, or have dual activity and are thus useful in the treatment of people who have immune allergic or cardiovascular problems caused by PAF or 5-lipoxygenase products.

B. Stcreochemie

Surprisingly, it has been determined that the activity and properties of the active compounds, including 5-lipoxygenase activity, oral availability and in vivo stability (e.g., glucuronidation ratio) may vary substantially in the disclosed compounds when switching from one optical isomer to another. Therefore, in a preferred embodiment, the active compound or precursor thereof is administered in enantiomerically enriched form, i.e. more or less in the form of a single isomer. The preferred enantiomer is readily determined by evaluating the different possible enantiomers in selected biological assays, e.g. those described in detail herein.

The 2,5-disubstituted tetrahydrofurans, tetrahydrothiophenes and pyrrolidines have a number of configurations. The carbon atoms 2 and 5 of the middle ring are chiral and so the middle ring exists at least as a diastereomeric pair. Each diastereomer exists as a series of enantiomers. Therefore, a compound based on chiral C2 and C5 atoms is a mixture of four enantiomers. If non-hydrogen substituents are present on the carbon atoms 3 and 4 of the middle ring, then the C3 and C4 atoms are also chiral, and therefore the ring exists as a diastereomeric pair and the compound is also a mixture of the four enantiomers.

The 1,3-cyclopentanes disclosed herein also occur in a variety of stereochemical configurations. The middle ring carbon atoms 1 and 3 are chiral and therefore the middle ring exists at least as a diastereomeric pair. Each diastereomer exists in a number of enantiomers. Therefore, a compound based on chiral C1 and C2 atoms is a mixture of four enantiomers.

When non-hydrogen substituents are present on the carbon atoms 3 and 4 of the middle ring, the C4 and C5 atoms are also chiral and the ring may exist as a diastereomeric pair, and the compound is a mixture of the four enantiomers.

One of ordinary skill in the art can readily prepare enantiomers of the disclosed compounds using chiral reagents and known procedures, and can evaluate the biological activity of the isolated enantiomers using the methods described herein or methods otherwise known. Optical enrichment can be determined by chiral NMR shift reagents, polarimetry or chiral HPLC.

Classical discrimination methods include various physical and chemical techniques. Often repeated recrystallization is the simplest and most effective technique. The recrystallization can be carried out at any stage of the preparation of the compound or in the case of the final enantiomeric product. If successful, it represents a unique method

If recrystallization does not give a substance of acceptable optical quality, other methods may be used. If the compound is basic, a chiral acid can be used to form diastereomeric derivatives which may have different dissolution properties. Some, but not limited, examples of chiral acids include malic acid, mandelic acid, dibenzoylsuccinic acid, 3-bromocapro-8-sulfonic acid, 10-camphorsulfonic acid and di-p-toluylsuccinic acid. Similarly, acylation of the free hydroxyl group with chiral acids leads to diastereomeric derivatives, whose physical properties may vary so important that separation can be performed.

Enantiometrically pure or enriched compounds can also be obtained by passing the racemic mixture through a chromatographic column which is intended for chiral separations or by enzymatic resolution of suitably modified substrates.

C. Synthesis of Active Compounds

The 2,5-disubstituted tetrahydrofurans, tetrahydrothiophenes and pyrrolidines disclosed herein can be prepared by a variety of methods known to those skilled in the art, including those disclosed by Whittaker, et al., Synlett, 1993 pp 111, Biorg. Copper. Lett., 1993 pp. 1499; Achiwa, et al., Chem. Pharm. Bull., 1989, pp. 1969.

For example, one method for the synthesis of enantiomerically enriched materials is presented in Scheme I. In this method, enantiometric synthesis begins with chiral reduction of ketones. After ring closure and reaction of the -OH group, the cis and trans isomers can be resolved by standard methods known to those skilled in the art, affecting diastereomeric resolution. Other chiral centers can be resolved by techniques known to those skilled in the art, including those illustrated below.

 ** * * á • · «« *

1,3-Disubstituted cyclopentanes can be prepared according to the procedure of Graham, et al. (1,3-diarylcyclopentanes: a new class of potent receptor antagonists. 197 ^th ACS National Meeting, Dallas, TX, April 19-14, 1989, Medical Chemistry Section, poster # 25 (abstract)) or other known methods.

The general procedure for the preparation of hydroxyurea is illustrated in Scheme 1 below:

_FL-NH2 triphosgene

R-N «C» O

R'NH (OH) .HCl

OH H ff C ^X

AND

O

Scheme 1 Preparation of hydroxyureas

General procedures for the preparation of return urea are illustrated in Scheme 2:

R = NOj

Zn / HCl or 5% Rh / C

N ₂ H ₂

R = NHOH

RŤÍCO

OH H

O

H

R CHj

NH 3 OH

NOH

NHOH i

fl ^x C ^x CH ₃

RTíCO

O

R ^X = ^X CH,

Scheme 2 Preparation of Retro-Hydroxyurea * a * a * a * a * a * a * a * a * a * a

• a a ·

The general procedure for the preparation of hydroxamic acid is presented in Scheme 3:

R = CO ₂ h oxaloyl chloride

R-COCl

RNH (OH) .HCl

OH

R * C ^X

Scheme 3 Preparation of hydroxamic acids

The general procedure for the preparation of retro-hydroxamic acids is presented in Scheme 4:

R = NHOH

RCOC1

TEA

OC (O) Rfi!

R ^{x X} C ^X HO

LiOH

OH

R *

R ^XX C ^X

Scheme 4 Preparation of retro-hydroxamic acids

Scheme 5 represents the synthesis of 2- (3,4,5-trimethoxyphenyl) -5- [3- (N'-substituted-N'-hydroxyureidyl) propoxy] tetrahydrofuran (1-4) and 2- (4-fluorophenyl) -5- [ 3- (N-substituted-N'-hydroxyureidyl) propoxy] tetrahydrofuran (9-12):

R | 'Rj' RjOCHj; ot Rj «F O., COO—

NiBH ₄ . THF, CH ₃ OH, H ₃ O -)

25 ° C4h

5M ŘiOŘUL

102 RcRi-Rj-OCH,

120 R? R? -H, R? F

trans isomers; 109 R ₍ · Rj-Rj-OCHj; 126 Rj * F cis isomers; 110 R _{t "} Rj" Rj "OCHj; 127 R _t " Rj "H.Ri" F

trans isomers ^: 1 R 1 · R 3 «R 3 · OCH 3, R 4 · CH 3

Ri R ₂ R *, -OCH _j4 R ^ -CHjCH HjCH,

R 1, R 3, H, R 3, R 4 -CH 3

R ₁ -R 1 -H, R 1 -F, 1 -CH 3 CH 3 CH 3 CH 3 cis isomers; 3 Rj R Rj R R, -OCHj, R ₄ CH CH,

Scheme 5 4 Rj-Rj-Rj-OCHj. R ^HjCHjCHjCHj

Ri «R]« H. Ri «F. _4-CH3

- 42'R, -R, -and Rj-F. R 6 -CH 2 CH 2 CH ₂ CH 3 shows the synthesis of 2- (2,4,5-trimethoxyphenyl) -5- (3hydroxyureidylpropoxy) tetrahydrofuran (13) and 2- (4-fluorophenyl) -5- (3hydroxyureidylpropoxy) tetrahydrofuran (14.15) ··· ♦ »1 * · 4 • **» «« · ♦

^ '^^^' Βτ

Rj

103 R @ 1 -R @ 1 -R @ 1 -OC1 R @ 123 R @ 1 --R @ 1 -R @ 3 RfF

O

KjCO ,. OMF

W ^K - * o

70 ° C 67 »

HIM'

106

HIM

Br

CO (CF3CO4CO, Β, Κ CHA 0 ° C30miA, 25 ° C 2h

DMFU;

trans isomers - 12 (R 1 -R _t (R 3 OCH)):

R 129, -R, -H.Rj-F cis isomers: 130 _t R «R« H. Rj-F trans isomers: 131 R 1 -R 1 -R 1 -OI 1;

132 Rt-Rj-LiR-F cis isomer: 133 Ri-Rj-ft, Rj-F

O-C-N-SKCH)

-AND

CHA, xyS / o ^from V'N ^from * 'wi. <“» «1 trans isomers: 134 R ^ RjaRjaOCHj:

135 R, -R ₁ H, R ₁ -F cis isomers: 136 R ₁ -R ₃ -H. Rj-F

^ ’^ / ^ Ν ^ ΝΗι

OH trans isomer: 13 R @ 1 -R @ 1 -R @ 1 CH 2;

R, -R, -H, Rj-F cis isomers: 15 RpRj-H / Rj-F »Scheme 6

• * » * * * * «· • # (r * * » * · * · • »» * · ··· • • « ♦ * • • • «» · · • · <· • · ♦ ·

Scheme 7 illustrates the synthesis of 2- _(3} 4-dimethoxyphenyl) -5- [3-N'-substituted-propoxy N'hydroxyureidyl jtetrahydrofuran (5-8):

115

O

116 -

R «a-Bu (4)

R »Me (5)

Scheme 7 · 4 4 4 4 · 4 4 4 4 4 «« «« «« «« «« «« «« «« «« «« «« 99 * ♦ 99 · · 99 99 99 99 99 99. 99 99 99

The following examples are illustrative only and are not intended to limit the entire scope of the invention.

Example 1 Preparation of 2- (3,4,5-trimethoxyphenyl) -5- [3- (N'-substituted-N'hydroxyureidyl) propoxy] tetrahydrofuran (1-4) and 2- (4-fluorophenyl) -5 * [ 3- (N'-substituted-N'-hydroxyuridyl) propoxy] tetrahydrofuran (912) (a) Preparation of 4- (3,4,5-trimethoxyphenyl) -4-one butyric acid t-butyl ester (compound 101)

3,4,5-Trimethoxybenzaldehyde (8.0 g, 40.77 mmol), tert-butyl acrylate (5.29 g,

41.29 mmol) and 3-ethyl-5- (2-hydroxyethyl) -4-methylthiazolium bromide catalyst (3.52 g, 13.95 mmol) were dissolved in 50 mL dimethylformamide (DMF). To this solution was added 5.86 mL of triethylamine. The reaction mixture was stirred at 60 ° C for 16 hours, cooled to room temperature and quenched by addition. 10% HCl (pH = 1-2) and extracted with dichloromethane. The organic layer was washed with water and saturated NaCl solution, dried over MgSO 4, filtered and evaporated in vacuo to an oil. The product was purified by column chromatography (silica, 3: 1 hexane / ethyl acetate) (4.5 g, 34%). 1 H NMR (CDCl 3): 1.46 (2.9H); 2.70 (t, 2 H); 3.24 (t, 2 H); 3.92 (s, 9H); 7.25 (s, 2 H).

(b) Preparation of 4- (4-fluorophenyl) -4-one-butyric acid t-butyl ester (compound 119) a This compound was prepared in a similar manner to that described in Example 1 (a), wherein 3,4,5-trimethoxybenzaldehyde with 4-fluorobenzaldehyde. 1 H NMR (CDCl ₃ ); 1.45 (s, 9H); 2.70 (t, 2 H); 3.23 (t, 2 H); 7.12 (m. 2H); 8.02 (m, 2 H).

(c) Preparation of 4- (3,4,5-trimethoxyphenyl) -4-hydroxybutyric acid t-butyl ester (Compound 102)

Ketoester 101 (1.09 g, 3.36 mmol) was added to 10 mL of THF and 20 mL of methanol.

To this reaction mixture was stirred at room temperature for 4 hours, quenched with water and extracted with ethyl acetate. The organic layer was washed with water, saturated with solution

NaCl, dried over MgSO 4, filtered, and concentrated in vacuo to afford the title product (1.13 g, 103%). 1 H NMR (CDCl ₃ ): 1.44 (s, 9H); 2.00 (m. 2H); 2.32 (m, 2 H); 4.72 (m, 1H); 7.01 (m, 2 H); 7.30 (m, 2 H).

(d) Preparation of 4- (4-fluorophenyl) -4-hydroxybutyric acid t-butyl ester (Compound 120)

This compound was prepared from 119 using the procedure outlined in Example 1 (c), wherein compound 101 was replaced with compound 119 ^. NMR (CDCl ₃ ): 1.44 (s, 9H); 2.00 (m. 2H); 2.32 (m, 2 H); 4.72 (m, 1H); 7.01 (m, 2 H); 7.30 (m, 2 H).

(e) Preparation of 4- (3,4,5-trimethoxyphenyl) -5-lactone (compound 104)

Hydroxyester 102 (1.13 g, 3.47 mmol) was added to 4 mL of methanol, 1.5 mL of water, and 5 M aqueous sodium hydroxide solution (4.5 mL). The reaction mixture was stirred at room temperature for 30 minutes and then 12 mL of saturated aqueous NaHCO 3 was added. The aqueous phase was washed with ether, acidified to pH 1-2 by addition of concentrated HCl and extracted with benzene (2 x 30 mL). The benzene layer was checked by TLC to show that a portion of the lactone had already formed. PPTS (10 mg) was added to the benzene extract and the mixture was refluxed for 1 hour to remove water. The reaction mixture was washed with saturated NaHCO ₃ solution and evaporated in vacuo to give the desired lactone as a solid (700 mg, 80%). ¹ H NMR (CDCl 3): 2.20 (m, 1H); 2.68 (m, 3H); 3.85 (s, 3H); 3.88 (s, 6H); 5.46 (m, 1H); 6.55 (s. 2H).

(f) Preparation of 4- (4-fluorophenyl) -3-lactone (Compound 102)

This compound was prepared from 120 using a procedure similar to that illustrated in Example 1 (e), wherein compound 102 was replaced by compound 120. 1 H NMR (CDCl 3): 2.20 (m, 1H); 2.68 (m, 3H); 5.50 (m, 1H); 7.10 (t, 2 H); 7.32 (m. 2H).

(g) Preparation of 2- (3,4,5-trimethoxyphenyl) -5-hydroxytetrahydrofuran (105)

Lactone 104 (6.86 g, 27.22 mmol) was dissolved in dry toluene (100 mL) and the solution was cooled to -70 ° C. DIBALH 1.5 M toluene solution (28 mL) was added dropwise to this solution. The reaction mixture was stirred at -70 ^° C for one hour. The reaction was slowed by the addition of methanol (11 mL) while the temperature was

ΛΛ,

'27 li C ί: tr r o r.

i: t:

b '<?

f ϋ fl »;

TO «Ii; efci r « «: £ i: * r f F,. Ť í · <

Network |, 4.

ø · '* ø ·' <'

4> F .i _Λ maintained at <-60 ° C. The mixture was warmed to -20 ° C, followed by the addition of a saturated aqueous solution of sodium potassium tartrate (96 mL), and the reaction temperature was maintained between * -10 ° C and 0 ° C. The reaction mixture was stirred at 0 ° C for 3 hours and then the two phases were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, saturated NaCl solution and concentrated in vacuo to give the product (6.51 g, 94%). 1 H NMR (CDCl 3): 1.82-2.48 (m, 4H); 3.84 (s, 3H); 3.88 (s, 6H); 4.97, 5.20 (m, 1H); 5.65, 5.79 (m, 1H); 6.56, 6.70 (s, 2H).

(h) - Preparation of 2- (4-fluorophenyl) -5-hydroxy tetrahydrofuran (123);

. ' 1. This compound was prepared from 122 using a procedure similar to Example 1> (g), substituting Compound 104 for compound 104. 1 H NMR (CDCl 3): 1.79 (m, 1H); 1-, 95-2.10 (m, 1H); 2.20-2.32 (m, 1H); 2.48 (m, 1H); 5.00 & 5.22 (m, 1H); 5.63 & 5.78 (m, 1H); 7.04 (m, 2 H); 7.30 & 7.41 (m, 2H).

(i) - Preparation of trans- and cis-2- (3,4,5-trimethoxyphenyl) -5- (3-phthalimidylpropoxy) tetrahydrofuran (compounds 107,108)

Compound 105 (1.14 g, 4.49 mmol) was dissolved in 4 mL of dichloromethane.

Triethylamine (681.4 mg, 6.73 mmol) was added to this solution. The reaction mixture was cooled in an ice bath and trifluoroacetic anhydride (1.41 g, 6.73 mmol) was added dropwise. The reaction mixture was stirred at 0 ° C for 30 minutes and then 3-phthalimidylpropanol (106) (2.4 g, 13.26 mmol) was added. The reaction mixture was warmed to room temperature and stirred at room temperature for 2 hours. The reaction was slowed with saturated aqueous NaHCO 3 and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated NaCl solution, dried over MgSO 4, filtered and evaporated in vacuo to an oil which was purified _by column chromatography (silica, 2: 1 hexane / ethyl acetate) (107: 522 mg (trans); 108;

271 mg (cis); mixture 1: 1107 and 108: 110 mg; overall yield 46%); ^! 1 H NMR (CDCl 3): 107: 1.70 (m, 1H); 1.82 (m, 1H); 2.00 (m. 2H); 2.02 (s, 1H); 2.28 (m, 1H); 3.46 (m, 1H); 3.83 (s, 3H); 3.84 (m, 3H); 3.88 (s, 6H); 4.99 (t, IH); 5.30 (dd, IH); 6.56 (s, 2 H); 7.72 (m, 2 H); 7.85 (m, 2 H). 108: 1.95 (m. 3H); 2.00 (m. 2H); 2.20 (m, 1H); 3.51 (m, 1H); 3.83 (s, 3H); 3.85 (m, 2 H); 3.88 (s, 6H); 3.92 (m, 1H); 4.90 (m, 1H); 5.16 (dd, 1H); 6.60 (s, 2 H); 7.72 (m, 2 H); 7.84 (m, 2 H).

· · * * * * * * * * * * * * * * * *

To determine the stereochemistry of this molecule, a NOE differential experiment was performed.

Trans (107) isomer: in this experiment, a triplet was irradiated at 4.99 ppm with a low-frequency spin disconnect pulse and the data was processed to merely measure the presence of an increase in signal. This represents a positive NOE effect and implies a close spatial relationship of these protons. In this experiment, NOE was found at 2.25-2.36 ppm, which represents furan protons. Another NOE was observed for the aromatic protons, indicating that this triplet was a benzylic proton. No NOE was observed for the double doublet at 5.30 ppm, suggesting it is a trans isomer.

Isomer cis (108): To determine stereochemistry in this molecule, a NOE differential experiment was performed. In this experiment, a multiplet was irradiated at 4.88-4.93 ppm with a low-frequency rf pulse disconnection pulse and the data was processed to only the presence of signal increase was measured. This represents a positive NOE effect and implies a close spatial relationship of these protons. In this experiment, a NOE for doublet was found at 5.16 ppm. which represents another methine proton. Another NOE was observed for aromatic protons, suggesting that this triplet is a benzylic proton. Additional NOE was observed for the multiplet at 1.93-2.20 ppm, which are additional furan methylene protons.

(j) Preparation of 2- (4-fluorophenyl) -5- (3-phthalimidylpropoxy) tetrahydrofuran (compounds 124,125)

These compounds were prepared from 123 by a procedure similar to that illustrated in Example 1 (i), substituting Compound 105 for Compound 105. 1 H NMR (CDCl ₃ ): 124 (trans): 1.65 (m, 1H); 1.80 (m, 1H); 2.00 (m. 2H); 2.12 (m, 1H); 2.31 (m, 1H); 3.48 (m, 1H); 3.82 (m, 3H); 3.82 (m, 3H); 5.02 (1H); 5.28 (dd, 1H); 7.00 (7.2H);

7.29 (m, 1H); 7.71 (m, 2 H); 7.85 (m, 2 H). 125 (cis): 1.90 (m, 2H); 1.99 (m, 4H); 2.19 (m, 1H); 3.48 (m, 1H); 3.82 (m, 2 H); 3.88 (m, 1H); 4.94 (m, 1H); 5.15 (dd, 1H); 7.00 (t. 2H); 7.30 (m, 2 H); 7.71 (m, 2 H); 7.84 (m, 2 H).

(k) Preparation of 3-phthalimidylpropanol (Compound 106) * in &quot; &quot; · I I I I I I I I I I I I I

3-Bromopropanol (4.0 g, 28.78 mmol), potassium phthalimide (8.0 g, 43.17 mmol) and potassium carbonate (4.0.28.78 mmol) were added to 20 mL of DMF. The reaction mixture was stirred at 70 ° C for 4 hours, quenched with water and extracted with ethyl acetate. The organic layer was washed with water, saturated NaCl solution and evaporated in vacuo to a solid which was crystallized in ethyl acetate (3.5 g, 67%).

(1) Preparation of trans- and cis-2- (3,4,5-trimethoxyphenyl) -5- (3aminopropoxy) tetrahydrofuran (compound 109,110)

Compound 107 (455 mg, 1.03 mmol) and hydrazine monohydrate (165.3 mg, 5.16 mmol) were added to 2 mL ethanol. The reaction mixture was refluxed for 2 hours, quenched with water and extracted with dichloromethane. The organic layer was washed with water and saturated NaCl solution, dried over MgSO ₄ , filtered and evaporated in vacuo to afford trans-109 (225 mg, 70%). 1 H NMR (CDCl ₃ ): 1.75 (m, 2H); 1.78 (m, 1H); 1.96 (m, 1H); 2.20 (m, 1H); 2.40 (m, 1H); 2.82 (t, 2 H); 3.55 (m, 1H); 3.81 (m, 1H); 3.83 (s, 3H); 3.87 (s, 6H); 5.00 (t, 1H); 5.34 (dd, 1H); 6.56 (s, 2 H).

The cis -isomer 110 was prepared from 108 by a procedure similar to that described in 109. 1 H NMR (CDCl ₃ ): 1.76 (m, 2H); 2.08 (m, 3H); 2.27 (m, 1H); 2.82 (t, 2 H); 3.55 (m, 1H); 3.8-4 (s, 3H); 3.88 (s, 6H); 3.92 (m, 1H); 4.95 (m, 1H); 5.20 (m, 1H); 6.64 (s, 2 H).

(m) Preparation of 2- (4-fluorophenyl) -5- (3-aminopropoxy) tetrahydrofuran (compounds 126 and 127)

These compounds were prepared from 124 and 125 by a procedure similar to that described in Example 1 (1), substituting compounds 107 and 108 for compounds 124 and 125. 1 H NMR (CDCl ₃ ): 124 (trans): 1.75 ( m, 3H); 1.96 (m, 1H); 2.20 (m, 1H); 2.40 (m, 1H); 2.82 (t, 2 H); 3.54 (m, 1H); 3.83 (m, 1H); 5.05 (t, 1H); 5.32 (dd, 1H); 7.01 (t, 2 H);

7.30 (m, 2 H). 125 (cis): 1.74 (m, 2 H); 1.97 (m, 1H); 2.05 (m, 2 H); 2.25 (m, 1H); 2.77 (t, 2 H); 3.47 (m, 1H); 3.85 (m, 1H); 4.95 (m, 1H); 5.15 (dd, 1H); 7.00 (t. 2H); 7.34 (m, 2 H).

(n) Preparation of trans- and cis-2- (3,4,5-trimethoxyphenyl) -5- [3- (N'-methyl-N'hydroxyureidyl) propoxy] tetrahydrofuran (compounds 1,3)

Compound 109 (60 mg, 0.19 mmol) and triphosgene (23 mg, 0.078 mmol) were dissolved in 3 mL of dichloromethane. To this solution was added triethylamine (29.3 mg, 0.29 mmol). The reaction mixture was refluxed for 2 hours and then cooled to b! p! fc.ti or t. pl t ct C Cí fitt frr *. rt c frrj

Ϊ “fc 11 V e t t tc« c. e t t Φ frt t

t ·

C.

ice bath. Triethylamine (34.0 mg, 0.34 mmol) and methylhydroxyamine hydrogen chloride (32.2 mg, 0.39 mmol) were added to the cold solution. The reaction mixture was stirred at ambient temperature for 16 hours, slowed with water and extracted with dichloromethane. The organic layer was washed with saturated NaCl solution and evaporated in vacuo to an oil which was purified by preparative TLC (silica, ethyl acetate) to give the product of trans compound 1 (51 mg, 69%). ¹ H NMR (CDCl 3): 1.82 (m, 3H); 1.95 (m, 1H); 2.22 (m, 1H); 2.40 (m, 1H); 3.15 (s. 3H); 3.40 > (m, 2H); 3.58 (m, 1H); 3.84 (s, 3H); 3.85 (m, 1H); 3.88 (s, 6H); 5.00 (t, 1H); 5.33 (m, 1H); 6.32 (m, 1H); 6.56 (s, 2 H); 7.37 (s, 1H). _{; 1>} - y / _{k, t} Isomer cis compound 3 was prepared from compound 110 using a procedure similar to that described for l.jHNMR _(CDCl3): 1.83 (m, 2H) ;. 2.07 ( m, 3H); 2.28 (m, 1H); 3.13 (s, 3H); 3.35 (m, 2 H); 3.55 µm, 1H); 3.84 (s, 3H); 3.87 (s, 6H); 3.88 (m, 1H); 4.97 (m, 1H); 5.20 (m, 1H); 6.22 (m, 1H); 6.63 (s, 2 H); 7.37 (s, 1H).

. (o) Preparation of 2- (4-fluorophenyl) -5- [3-N'-methyl-N'-hydroxyureidyl) propoxy) tetrahydrofuran (compounds 9.11) ¹

These compounds were prepared from. 126 and 127 using the procedure described in Example 1 (n) wherein compounds 109 and 110 were replaced with compounds. 126 and 127. 1 H NMR (CDCl 3): δ (trans): 1.70 (m, 1H); Δ, 78 (m, 2H); L, 96 (m, 1H); 2.19 (m, 1H); 2.40 (m, 1H); 3.10 (s, 3H); 3.31 (m, 2 H); 3.51 (m, 1H); 3.83 (m, 1H); 5.05 (t, 1H); 5.30 (dd, 1H); 6.38 (t, 1H); 7.01 (t, 2 H); 7.28 (m, 2 H). Δ (cis): 1.80 (m, 2H); 2.05 (m. 3H); 2.24 (m, 1H); 3.06 (s, 3H); 3.30 (m, 2 H); 3.48 (m, 1H); 3.86 (m, 1H); 4.98 (m, 1H); 5.16 (dd, H); 6.30 (t, 1H); 7.02 (t, 1H); 7.31 (m, 2 H); 8.08 (bs, 1H). ,> ..

(p) Preparation of trans- and cis-2- (3,4,5-tripiethoxyphenyl) -5- [3- (N'-n-butyl-N'hydroxyureidyl) propoxy] tetrahydrofuran (compound 2,4), Compound 109 ( 60 mg, 0 ₁₉ mmol) and triphosgene (23 mg, 0.078 mmol) were dissolved in 3 ml of dichloromethane. To this solution was added triethylamine (29.3 mg, 0.29 mmol). The reaction mixture was refluxed for 2 hours and then cooled in a water bath. Butyl hydroxylamine (51.4 mg, 0.29 mmol) was added to the cold solution. The reaction mixture was stirred at room temperature for 16 hours, cooled with water and extracted with dichloromethane. The organic layer was washed with a saturated solution of 9 ° C and 9 ° C. · · · · ·

NaCl and evaporated in vacuo to an oil. The trans product of compound 2 was separated by preparative TLC (silica, ethyl acetate) (46.9 mg, 57%). 1 H NMR (CDCl 3): 0.93 (t, 3H); 1.35 (m, 2 H); 1.58 (m, 2 H); 1.81 (m, 3H); 1.96 (m, 1H); 2.21 (m, 1H); 2.40 (m, 1H); 3.38 (m, 2 H); 3.50 (m, 2 H); 3.57 (m, 1H); 3.83 (s, 3H); 3.85 (m, 1H); 3.88 (s, 6H); 5.00 (t, 1H); 5.32 (m, 1H); 6.32 (m, 1H); 6.56 (s, 2 H).

The cis isomer of compound 4 was prepared from 110 using a procedure similar to that described for compound 2. 1 H NMR (CDCl 3): 0.92 (t, 3H); 1.32 (m, 2 H); 1.58 (m, 2 H); 1.81 (m, 2 H); 2.08 (m, 3H); 2.28 (m, 1H); 3.35 (m, 2 H); 3.47 (m, 2 H); 3.54 (m, 1H); 3.84 (s, 3H); 3.87 (s, 6H); 3.88 (m, 1H); 4.97 (m, 1H); 5.20 (m, 1H); 6.22 (m, 1H); 6.63 (s, 2 H).

(q) Preparation of 2- (4-fluorophenyl) -5 [3- (N'-n-butyl-N'-hydroxyureidyl) propoxy] tetrahydrofuran (compounds 10,12)

These compounds were prepared from 126 and 127 using the procedure described in Example 1 (p); wherein compounds 109 and 110 were replaced by compounds 126 and 127. 1 H NMR (CDCl 3): 10 (trans): 0.90 (t, 3H); 1.30 (m, 2 H); 1.55 (m, 2 H); 1.70 (m, 1H); 1.78 (m, 2 H); 1.96 (m, 1H); 2.19 (m, 1H); 2.40 (m, 1H); 3.31 (m, 2 H); 3.44 (m, 2 H); 3.52 (m, 1H); 3.82 (m, 1H); 5.05 (t, 1H); 5.30 (dd, 1H); 6.32 (t, 1H); 7.00 (t. 2H); 7.28 (m, 2 H); 7.55 (bs, 1H). 12 (cis): 0.90 (t, 3H); 1.30 (m, 2 H); 1.52 (m, 2 H); 1.80 (m, 2 H); 2.04 (m, 3H); 2.24 (m, 1H); 3.30 (m, 2 H); 3.40 (m, 2 H); 3.48 (m, 1H); 3.85 (m, 1H); 4.98 (t, 1H); 5.16 (dd, 1H); 6.27 (t, 1H); 7.03 (t, 2 H); 7.32 (m, 2 H); 7.53 (bs, 1H).

Example 2 Preparation of 2- (3,4-dimethoxyphenyl) -5- [3-N ^, -substituted-N'hydroxyureidylpropoxy] tetrahydrofuran (5-8) (a) Preparation of 4- (3 ', 4'-dimethoxyphenyl) -4-oxobutyrnitrile (111)

One portion of pure acetonitrile (3.32 mL, 0.048 mol) and triethylamine (5 mL, 0.11 mol) were added to a stirred mixture of 3-benzyl-5- (2-hydroxyethyl) -4-methylthiazolium chloride (5.3 g). , 0.02 mol) in dry dimethylformamide (25 ml) under an argon atmosphere. The mixture was allowed to stand overnight at ambient temperature. The reaction mixture was diluted with water and extracted with ethyl acetate (3 × 100 mL). The organic extract was washed with water (3 x 100 mL), brine (3 x 100 mL) and the solvent was removed under reduced pressure to give an amber oil. TLC analysis (silica gel, 4 * 4) »4 * 4 * 4 * 4 * 4 * 4 Ethyl acetate: hexanes, 1: 1) showed the presence of a mixture of three spots at Rf 0.80 (starting aldehyde), 0.50 (Compound 1). ) and 0.30 (unknown by-product). The sample was purified by silica gel 60 column chromatography (230-400 mesh) and eluted with ethyl acetate: hexanes (1: 1) to give the title compound as a yellow product (2.26 g, 22%). 1 H NMR (CDCl ₃ ): 2.78 (t, 2H, J = 8Hz), 3.33 (t, 2H, J = 8Hz, 3.96 (s, 3H); 3.98 (s, 3H); 6.90 (d, 1H, J = 8.5 Hz, 7.52 (d, J = 2.2H); 7.58 (dd, J = 2 and 8 Hz, 2H).

(b) Preparation of 4- (3,4-dimethoxyphenyl) -4-oxo-butyric acid (112)

A stirred solution of 4- (3 ', 4'-dimethoxyphenyl) -4-oxobutyronitrile (111) (2.26 g, 0.01 mol) in acetic acid (15 mL) and hydrochloric acid (12 N, 40 mL) was heated. under reflux for 1.5 hours and cooled to room temperature. The solvent was removed under reduced pressure to give a brown solid. Recrystallization from water gave 112 as pale crystals (1.57 g, 66%). 1 H NMR (CDCl ₃ ) 2.80 (t, J = 7.5 Hz, 2H); 3.30 (t, J = 7.5Hz, 2H); 3.94 (s, 3H); 3.96 (s, 3H); 6.89 (d, 1H, J = 9Hz); 7.55 (d, 1H, J = 1 Hz) and 7.64 (dd, 1H, 1 and 9 Hz).

(c) Preparation of 4- (3 ', 4'-dimethoxyphenyl) butyrolactone (113)

A solution of sodium borohydride (0.89 g, 0.023 mol) in water (4 mL) was added dropwise (ca, 5 min) to a stirred solution of 112 (2.8 g, 0.022 mol) in freshly distilled tetrahydrofuran (40 mL). ) and methanol (20 mL) under an argon atmosphere. The reaction was allowed to stand overnight at room temperature. TLC analysis (silica gel, ethyl acetate: methanol: acetic acid, 9.5: 0.5: few drops) revealed the presence of starting material. An additional portion of sodium borohydride (0.5 g, 0.013 mol) in water (2 mL) was added dropwise and the reaction was allowed to stand for three hours. TLC analysis (same system as above) showed no starting material. The reaction was quenched with hydrochloric acid (6 N, 25 mL) and allowed to stand for 15 minutes. The mixture was extracted with ethyl acetate (3 x 75 mL). The organic extract was washed with water (3 x 75 mL), brine (3 x 75 mL), and the solvent was removed under reduced pressure to give a solid (2.0 g, 75%). 1 H NMR (CDCl ₃ ): 2.18-2.25 (m, 1H); 2.59-2.70 (m, 3H); 3.89 (s, 3H); 3.90 (s, 3H); 5.44-5.49 (m, 1H) and 6.82 -6.87 (m, 3H).

* ♦ ** * »99 · ♦» * * * * * 99 99 99 99 99

(D) Preparation of 4- (3 ', 4'-dimethoxyphenyl) butyrolactol

A solution of diisobutylaluminum hydride (1.5 M, 9 mL, 13.5 mmol) was added dropwise (ca. 30 min) to 113 (2.0 g, 9 mmol) in dry toluene (40 mL) under an argon atmosphere which was cooled in a dry ice-acetone bath. The reaction mixture was stirred at -78 ° C for one hour. TLC analysis (silica gel, ethyl acetate; hexanes, 1: 1) did not show the presence of starting material, but revealed the presence of a new spot in Rf 0.38. The reaction was quenched with methanol (20 mL) and the lesson warmed to 0 ° C. Saturated sodium potassium tartrate solution (50 mL) was then added and the mixture was stirred at 0 ° C for 45 minutes. The mixture was extracted with ethyl acetate (3 x 100 mL) and the organic extract was washed with water (3 x 75 mL) and brine (3 x 75 mL). Removal of the solvent under reduced pressure gave a dark amber oil (1.7 g, 84%). 1 H NMR (CDCl 3) (mixture of cis and trans isomers); 1.71 - 2.49 (m, 8H); 2.91 (br s, 1H); 3.09 (br s, 1H); 3.89 (s, 6H); 3.90 (s, 6H); 4.97 (m, IH); 5.19 (t, J = 7 Hz, 1H); 5.62 (m, 1H); 5.77 (m, 1H) and 6.82-7.28 (m, 6H).

(e) Preparation of N- (3-hydroxypropyl) phthalimide (106)

A mixture of 3-bromopropanol (4 g, 0.029 mol), potassium phthalate (8 g, 0.043 mol) and potassium carbonate (4 g, 0.029 mol) in dry DMF (50 mL) were stirred and heated at 70 ° C for four hours . The mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 x 75 mL). The organic extract was washed with water (3 x 100 mL) and dried (Na 2 SO ₄ ). Removal of the solvent was performed at low pressure to give a white solid which was recrystallized to give white crystals (1.27 g, 24%).

(f) Preparation of trans- and cis-2- (3 ', 4'-dimethoxyphenyl) -5- [3- (N-phthaloyl)] propoxy-tetrahydrofuran (115 and 116)

Triflic anhydride (0.68 mL, 4.8 mmol) was added in one portion to a stirred solution of 114 (0.72 g, 3.2 mmol) in dry dichloromethane (20 mL) and triethylamine (0.68 mL, 4, 0.5 mL). 9 mmol) under an argon atmosphere, the mixture was cooled using an ice bath. The reaction mixture was stirred at 0 ° C for 30 minutes. To the reaction mixture was added N (3-hydroxypropyl) phthalimide (106) (1.27 g, 7 mmol) and the solution was allowed to warm to room temperature and allowed to stand at this temperature for two hours. The solution was quenched with aqueous dodecarbonate solution (saturated, 25 mL) and extracted

• · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · Ethyl acetate (3 χ 50 ml), brine (3 × 50 mL) and dried (sodium sulfate). Removal of the solvent under reduced pressure gave an amber oil (2.02 g). TLC analysis of the oil (silica gel; ethyl acetate: heany, 1: 1) showed the presence of four spots at Rf 0.80; 0.60; 0.50 and 0.35. The spots at Rf 0.60 and 0.50 were 2: 1. The sample was purified by silica gel flash chromatography (230-400 mesh) and eluted with ethyl acetate: hexanes (3: 7) to give a clear and colorless oil with Rf 0.60 (0.40 g, 30%), this The product was identified as trans-2- (3 ', 4'-dimethoxyphenyl) -5 [3- (N-phthaloyl)] propoxytetrahydrofuran (115). (0.40 g, 30%). H NMR (CDCl3): 1.34 1.94 (m, 2H); 1.96 - 2.05 (m, 2H); 2.09 - 2.20 (m, 1H); 2.25-2.36 (m, 1H); 3.46-3.53 (m, 1H); 3.84 (t, 9Hz, 2H), there is also a hidden multiplet of one proton, 3.88 (s, 3H); 3.91 (s, 3H); 5.01 (t, 7.3 Hz, 1H); 5.30 (dd, J = 2 and 5 Hz, 1 Hz); 6.82 - 6.90 (m, 3H); 7.71 - 7.74 (m, 2H) and 7.84 - 7.88 (m, 2H).

To determine the stereochemistry of this molecule, a differential NOE experiment was performed. In this experiment, a triplet at 5.01 ppm would be irradiated with a low-frequency rf pulse to cancel the spin interaction, and the data was processed by merely measuring the presence of signal magnification. This represents a positive NOE effect, indicating the spatial presence of these protons. In this experiment, NOE was found at a multiplet of 2.25-2.36 ppm, which are furan protons. Another NOE was observed for aromatic protons, revealing the presence of benzylic protons. NOE for double doublet at 5.30 ppm was not observed, indicating that the substance is a trans isomer.

Continued elution with an identical solvent system gave a spot of Rf 0.50, a colorless oil (0.21.15%) was identified as cis- (2-3 ', 4'-dimethoxyphenyl) -5- [3- (N-phthaloyl)]. Propoxytetrahydrofuran (116). 1 H NMR (CDCl ₃ ): 1.92-2.12 (m, 6H); 3.44 - 3.52 (m, 1H); 3.86 (s, 3H); 3.76 - 3.93 (m, 3H); 4.89 - 4.94 (m, 1H); 5.35 (d, &gt; 4 Hz); 6.89 (d, J = 8Hz); 6.89 (dd, &gt; 2 and 8 Hz); 6.92 (d, J = 2H); 7.69 7.72 (m, 2H) and 7.82-7.85 (m, 2H).

To determine the stereochemistry of this molecule, a differential NOE experiment was performed. In this experiment, a multiplet at 4.89-4.94 ppm was irradiated with a low-frequency rf pulse canceling the spin interaction and data processing was performed by measuring only signal magnification. This represents a positive NOE effect that t

4 «» ♦ ·

· · · · · · · · · · · ·

· «*

· «Indicates the close spatial relationship of these protons. In this experiment, a NOE was found for a doublet with 5.35 ppm, which represents another methine proton of furan. This indicates that the molecule is the cis isomer. Additional NOE was observed with aromatic protons suggesting that this triplet was a benzylic proton. There is also another NOE for the 1.92-1.12 ppm multiplet, which contains non-methyl protons of furan.

Chromatography also gave a mixture of 115 and 116.

(g) Preparation of trans-2- (3 ', 4'-dimethoxyphenyl) -5- (3aminopropoxy) tetrahydofuran (117)

Pure hydrazine hydrate (150 μΐ, 3.2 mmol) was added to a stirred solution of 115 (253 mg, 0.62 mmol) in absolute ethanol (1.5 mL). The solution was refluxed for 5 minutes, whereupon a white solid precipitated. The mixture was heated under reflux for an additional two hours. TLC analysis (silica gel; ethyl acetate / hexanes, 1: 1) showed no starting material but a starting spot. The reaction mixture was quenched with water (10 mL) and extracted with dichloromethane (5x10 mL). The organic phase was washed with water (2 x 10 mL), brine (2 x 10 mL) and dried (sodium sulfate). Removal of the solvent under low pressure gave a colorless oil (150 mg, 86%). 1 H NMR (CDCl ₃ ) 1.25 (br s, 2H); 1.68-1.78 (m, 3H); 1.81 - 1.98 (m, 1H); 2,14,2,2 (m, 1H); 2.3-2.36 (m, 1H); 2.80 (t, J = 6.5Hz, 2H); 3.47-3.55 (m, 1H); 3.78 - 3.87 (m, partially hidden, 1H); 3.86 (s, 3H); 3.88 (s, 3H); 4.99 (t,> 7 Hz, 1H); 5.31 (dd, J = 2 and 6 Hz, 1H); 6.80-6.88 (m, 3H).

(h) Preparation of cis-2- (3 ', 4'-dimethoxyphenyl) -5- (3aminopropoxy) tetrahydrofuran (118)

Pure hydrazine hydrate (125 μΐ, 2.57 mmol) was added to a stirred solution of 116 (210 mg, 0.51 mmol) in absolute ethanol (3.0 mL). The solution was refluxed for 5 minutes, whereupon a white solid precipitated. The mixture was refluxed for two hours. TLC analysis (silica gel, ethyl acetate: hexanes, 1: 1) did not show the presence of the starting material but showed the presence of a starting spot. The reaction was quenched with water (10 mL). The organic phase was washed with water (2 x 10 mL), brine (1 x 10 mL) and dried (sodium sulfate). Removal of the solvent under reduced pressure gave a solid oil (150 mg, 73%). ^! 1 HNMR (CDCl ₃₎ • 9

• 99

9 «

9

9

9 * 9 * 99

99

9 9 9

9 99

999 9 · 9 9

99

1.45 (br s, 2H); 1.73-1.78 (m, 2H); 2.01-2.12 (m, 3H); 2.19 - 2.29 (m, 1H); 2.81 (t, J = 7Hz, 2H); 3.48 - 3.53 (m, 1H); 3.85 - 3.93 (m, partially hidden, 1H); 3.88 (s, 3H); 3.90 (s, 3H); 4.96 - 5.01 (m, 1H); 5.17 (dd, J = 3 and 6 Hz, 1H); 6.83 (d, J = 8Hz, 1H); 6.89 (dd, J = 2 and 8 Hz, 1H) and 6.96 (d, J = 2 Hz, 1H).

(i) Preparation of trans-2- (3 ', 4'-dimethoxyphenyl) -5- [3- (N-butyl-Nhydroxyureidyl) propoxy] tetrahydrofuran (5)

Triethylamine (32 μΐ, 0.22 mmol) and then triphosgene (19 mg, 0.06 mmol) were added to a stirred solution of 117 (53 mg, 0.19 mmol) in dry dichloromethane (3 mL) under an argon atmosphere. The solution was heated under reflux for 30 minutes and cooled to room temperature. Solid n-butylhydroxylamine (34 mg, 0.38 mmol) was added in one portion to this solution, which was then allowed to stand over at ambient temperature. The reaction was slowed with water (10 mL) and the mixture was extracted with dichloromethane (3 x 10 mL). The combined organic phases were washed with aqueous sodium bicarbonate solution (saturated, 3 x 10 mL) and dried (sodium sulfate). TLC analysis (silica gel, ethyl acetate) revealed a complex mixture with Rf 0.90; 0.50; 0.25 and 0.00. The sample was purified by silica gel 60 flash chromatography (mesh 230-240) and eluted with ethyl acetate to give a spot with Rf 0.50 as an opaque oil (8 mg, 11%). ^! 1 H NMR (CDCl 3): 0.92 (t,> 7 Hz, 3H); 1.27 - 1.39 (m, 2H); 1.51 - 1.61 (m, 2H); 1.71 - 1.86 (m, 3H); 1.88-2.15 (m, 1H); 2.17-2.29 (m, 1H); 2.32-2.42 (m, 1H); 3.28-3.58 (m, 4H); 3.81 - 3.94 (m, partially hidden, 2H); 3.87 (s, 3H); 3.90 (s, 3H); 5.49 - 5.05 (m, 1H); 5.31 - 5.38 (m, 1H); 6.28 - 6.34 (m, 1H) and 6.81 - 6.86 (m, 3H). IR (film); 3407,3193,2933,1640,1516,1263,1029 cm -1 ^! .

(j) Preparation of trans-2- (3 ', 4'-dimethoxyphenyl) -5- [3- (N-methyl-N-hydroxyuridyl) propoxy] tetrahydrofuran (6)

Trifosgene (12 mg, 0.04 mmol), immediately followed by triethylamine (17 µΐ, 0.12 mmol) was added to a stirred solution of 117 (32 mg, 0.011 mmol) in dry dichloromethane (3 mL) under an argon atmosphere. The solution was heated under reflux for 2 hours, cooled to room temperature and placed in an ice bath. Pure triethylamine (32 µL, 0.23 mmol) was added to the reaction mixture. The reaction was allowed to stand at room temperature overnight. It was then quenched with water (10 mL) and extracted

9 9 9 9 9 9 9 9 99 dichloromethane (3 x 10 ml). The organic extract was washed with water (3 x 10 mL), brine (3 x 10 mL) and the solvent was removed under reduced pressure to give an amber oil. TLC analysis (silica gel, ethyl acetate) revealed only one new spot with Rf 0.30. The sample was purified by silica gel 60 column chromatography (230-240 mesh) and eluted with ethyl acetate to provide the desired compound as an amber oil (12 mg, 30%). ¹ H NMR (CDCl 3): 1.73-1.84 (m, 2H); 1.90-2.01 (m, 1H); 2.03 - 2.13 (m, 1H); 2.18-2.29 (m, 1H); 2.32-2.43 (m, 1H); 3.13 (s, 3H); 3.30-3.44 (m, 2H); 3.49 - 3.59 (m, 1H); 3.82 - 3.92 (m, partially hidden, 3H); 3.88 (s, 3H); 3.91 (m, 3H); 4.96 - 5.04 (tn, 1H); 5.34 (dd, J = 2 and 5 Hz, 1H); 6.34 (br t, 5Hz, 1H) and 6.82-6.68 (m, 3H). IR (film): 3407, 2229, 2935, 1636, 1516, 1263 and 1029 cm ^-1 .

(k) Preparation of cis-2- (3 *, 4'-dimethoxyphenyl) -5- [3- (N-butyl-Nhydroxyureidyl) propoxy] tetrahydrofuran (8)

Trifosgene (18 mg, 0.06 mmol), immediately followed by triethylamine (80 µΐ, 0.57 mmol) was added to a stirred solution of 118 (50 mg, 0.18 mmol) in dry dichloromethane (3 mL) under an argon atmosphere. The solution was heated under reflux for 2 hours, cooled to room temperature and placed in an ice bath. Pure triethylamine (50 μΐ, 0.35 mmol) was added, followed by solid n-butylhydroxylamine (32 mg, 0.36 mmol). The reaction was allowed to stand overnight at room temperature. It was then quenched with water (10 mL) and extracted with dichloromethane (3 x 10 mL). The organic extract was washed with water (3 x 10 mL), brine (3 x 10 mL), and the solvent was removed under reduced pressure to give an amber oil. TLC analysis (silica gel, ethyl acetate) revealed two new spots in approximately equal amounts with Rf 0.85 and 0.45. The sample was purified by silica gel 60 flash column (230-400 count) and eluted with ethyl acetate to give a first spot with Rf 0.85 as an amber oil (26 mg). Continued elution with identical solvents gave the title compound as an amber oil (25 mg, 35%). ^! 1 H NMR (CDCl 3): 1.1 (t, J = 7 Hz, 3H); 1.25-1.37 (m, 2H); 1.49-1.59 (m, 2H); 1.76-1.84 (m, 2H); 1.99-2.1 (m, 3H); 2.19 - 2.26 (m, 1H); 3.26-3.54 (m, 5H); 3.84 - 3.92 (m, partially hidden, 1H); 3.87 (s, 3H); 3.88 (s, 3H); 4.94 - 5.02 (m, 1H); 5.17 (d, J = 4Hz,

• ** • · · · • • · • · * · · » • • · · · · * • · ·· « • · · • * • ·· ·· ·· ··· • » • *

IH); 6.24 (t, J = 4 Hz, 1H); 6.52 (br s, 1H); 6.83 (d,> 8 Hz, 1H) and 6.89-95 (m, 2H). IR (film): 2913, 1640, 1570, 1463, 1262, 1139, and 1031 cm ⁴ .

(1) Preparation of cis2- (3 ', 4'-diethoxyphenyl) -5- [3- (N-methyl-N-hydroxyureidyl) propoxy] tetrahydrofuran (8)

Trifosgene (20 mg, 0.07 mmol) immediately followed by triethylamine (80 μΐ, 0.57 mmol) was added to a stirred solution of 118 (56 mg, 0.2 mmol) in dry dichloromethane (3 mL) under an argon atmosphere. The solution was heated under reflux for 2 hours, cooled to room temperature and placed in an ice bath. Pure triethylamine (80 μΐ, 0.57 mmol) was added, followed by solid methylhydroxyammonium hydrogen chloride (32 mg, 0.39 mmol). The reaction was allowed to stand overnight at room temperature. It was then cooled with water (10 ml) and the mixture was extracted with dichloromethane (3 × 10 ml). The organic extract was washed with water (3 x 10 mL), brine (3 x 10 mL) and the solvent was removed under reduced pressure to give an amber oil. TLC analysis (silica gel, ethyl acetate) revealed a spot at rf of 0.30 and some anion at the start. The sample was purified by silica gel 60 column chromatography (mesh 230-400) and eluted with ethyl acetate to give the above compound as an amber oil (30 mg, 42%). ^! 1 H NMR (CDCl ₃ ): 1.76 (m, 2H); 1.98 (2.10) m, 3H); 2.18 - 2.26 (m, 1H); 3.07 (s, 3H); 3.25-3.37 (m, 2H); 3.46 - 3.54 (m,

IH); 3.85 - 3.90 (m, partially hidden, 1H); 3.87 (s, 3H); 3.88 (s, 3H); 4.93 - 5.00 (m,

1H); 5.16 (d, J = 4 Hz, 1H); 6.27 (t, J = 5 Hz, 1H); 6.83 (d, J = 8 Hz, 1H) and 6.88-6.93 (m, 2H). IR (neat) 2933, 1663, 1518, 1621 and 1029 cm ^-1 .

Example 3 Preparation of 2- (2,4,5-trimethoxyphenyl) -5- (3-hydroxyureidylpropoxy) tetrahydrofuran (13) and 2- (4-fluorophenyl) -5- (3-hydroxyureidylpropoxy) tetrahydrofuran (14,15) (a ) Preparation of 2- (3,4,5-trimethoxyphenyl) -5- (3-bromopropoxy) tetrahydrofuran (Compound 128)

Compound 105 (1.0 g, 3.94 mmol) was dissolved in 4 mL of dichloromethane.

To this solution was added triethylamine (597 mg, 5.90 mmol). The reaction mixture was cooled in an ice bath and trifluoroacetic anhydride (1.24 g, 5.90-4 mmol) was added dropwise. The reaction mixture was stirred at 0 ° C for 30 minutes and then 3-bromopropanol (1.84 g, 13.27 mmol) was added. The reaction mixture was warmed to room temperature and stirred at room temperature for 2 hours. The reaction was quenched with saturated NaHCO 3 solution and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated NaCl solution, dried over MgSO ₄ , filtered and evaporated in vacuo to an oil which was purified by column chromatography (silica, 4: 1 hexane / ethyl acetate) (128: 430 mg and cis-isomer). 250 mg, total yield 46%). NMR (CDCl 3): 128 (trans-): 1.77 (m, 1H); 1.98 (m, 1H); 2.15 (m, 1H); 2.20 (m, 1H); 2.40 (m, 1H); 3.53 (t, 2 H); 3.60 (m, 1H); 3.83 (s, 3H); 3.87 (m, 1H); 3.89 (s, 6H); 5.01 (t, 1H); 5.35 (dd, 1H); 6.57 (s, 2 H).

(b) Preparation of 2- (4-fluorophenyl) -5- (3-bromopropoxy) tetrahydrofuran (compounds 129,130)

These compounds were prepared from 123 using the procedure of Example 3 (a), substituting compound 105 for compound 105. 1 H NMR (CDCl 3): 129 (trans-): 1.72 (m, 1H); 1.98 (m, 1H); 2.14 (m, 2 H); 2.20 (m, 1H); 2.40 (m, 1H); 3.53 (t, 2 H); 3.60 (m, 1H); 3.89 (m, 1H); 5.06 (t, 1H); 5.34 (m, 1H); 7.02 (t, 2 H); 7.30 (m, 2 H). 130 (cis -): 1.98 (m, 1H); 2.07 (m, 2 H); 2.14 (m, 2 H); 2.26 (m, 1H); 3.52 (t, 2 H); 3.58 (m, 1H); 3.93 (m, 1H); 5.00 (m, 1H); 5.20 (dd, 1H); 7.03 (t, 2 H); 7.35 (m, 2 H).

(c) Preparation of 2- (3,4,5-trimethoxyphenyl) -5- (3-Obenzylhydroxylaminopropoxy) tetrahydrofuran (Compound 131)

Compound 128 (260 mg, 0.69 mmol) was dissolved in 2 mL of 1,3-dimethyl-3,4,5,6-tetrahydro-2- (1H) -pyrimidone (DMPU). To this solution was added sodium carbonate (220.4 mg, 2.08 mmol) and benzylhydroxylamine hydrogen chloride (166 mg, 1.04 mmol). The reaction was stirred at 80 ° C for 16 hours, cooled with water and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over MgSO ₄ , filtered, and evaporated to an oil which was purified by flash column chromatography using ethyl acetate (114 mg, 40%). ¹ H NMR (CDCl 3): 1.72 (m, 1H); 2.36 (m, 1H); 3.06 (t, 2 H); 3.52 (m, 1H); 3.81 (m, 1H); 3.83 (s, 3H); 3.87 (s, 6H); 4.71 (s, 2 H); 4.98 (t, 1H); 5.30 (dd, 1H); 6.55 (s, 2 H); 7.35 (m, 5H).

· · V v v v v v v v v v v v v v v v v v v v v v v v v v v v v v v (D) Preparation of 2- (4-fluorophenyl) -5- (3-O-benzylhydroxylaminopropoxy) tetrahydrofuran (compounds 132, 133)

These compounds were prepared from compounds 129 and 130 using a procedure similar to Example 3 (c), wherein compound 128 was compound 129 and 130 ^. 1 H NMR (CDCl ₃ ): 132 (trans-): 1.70 (m, 1H); 1.83 (m, 2 H); 1.94 (m, 1H); 2.17 (m, 1H); 2.38 (m, 1H); 3.07 (t, 2 H); 3.52 (m, 1H); 3.82 (m, 2 H); 4.71 (s, 2 H); 5.02 (t, 1H); 5.30 (ss, 1H); 7.02 (t, 2 H); 7.30 (m, 2 H); 7.36 (m, 5H). 133 (cis): 1.85 (m, 2H); 1.96 (m, 1H); 2.05 (m, 2 H); 2.26 (m, 1H); 3.05 (t, 2 H); 3.50 (m, 1H); 3.88 (m, 2 H); 4.70 (s, 2 H); 4.99 (m, 1H); 5.17 (dd, 1H); 5.50 (bs, 1H); 7.00 (t. 2H); 7.35 (m, 7H).

(e) Preparation of 2- (3,4,5-trimethoxyphenyl) -5- (3-benzylhydroxyureidylpropoxy) tetrahydrofuran (Compound 134)

Compound 131 (114 mg, 0.27 mmol) was dissolved in 3 mL of dichloromethane.

To this solution was added trimethylsilyl isocyanate (47.6 mg, 0.41 mmol). The reaction mixture was stirred at ambient temperature for 16 hours and then refluxed for 4 hours. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate and evaporated to an oil. The product was isolated by preparative TLC using ethyl acetate as solvent. 1 H NMR (CDCl ₃ ): 1.72 (m, 1H); 1.94 (m, 3H); 2.16 (m, 1H); 2.38 (m, 1H); 3.50 (m, 1H); 3.62 (m, 2 H); 3.80 (m, 1H); 3.82 (s,

3H); 3.84 (s, 6H); 4.81 (s, 2 H); 4.99 (t, 1H); 5.30 (m, 3H); 6.54 (s, 2 H); 7.37 (s, 5H).

(f) Preparation of 2- (4-fluorophenyl) -5- (3-O-benzylhydroxyureidylpropoxy) tetrahydrofuran (compounds 135, 136)

These compounds were prepared from 132 and 133 using the procedure of Example 3 (e), substituting Compound 131 for Compounds 132 and 133. NMR (CDCl ₃ ): 135 (trans): 1.70 (m, 1H); 1.93 (m, 3H); 2.16 (m, 1H); 2.39 (m, 1H); 3.50 (m, IH); 3.62 (m, 2 H); 3.80 (m, 1H); 4.82 (s, 2 H); 5.04 (t, 1H); 5.30 (dd, 1H); 5.35 (bs, 2 H); 7.00 (t. 2H); 7.29 (m, 2 H); 7.38 (s, 5H). 136 (cis): 1.98 (m, 4H); 2.08 (m, 1H); 2.25 (m, 1H); 3.48 (m, 1H); 3.62 (m, 2 H); 3.83 (m, 1H); 4.81 (s, 2 H); 4.98 (m, 1H); 5.17 (dd, 1H); 5.42 (bs, 1H); 7.00 (t. 2H); 7.33 (m, 2 H); 7.38 (s, 5H).

(g) Preparation of 2- (3,4,5-trimethoxyphenyl) -5- (3-hydroxyureidypropoxy) tetrahydrofuran (Compound 13)

· * * * * * * * * * * * * * *

Compound 134 (90 mg, 0.19 mmol) was dissolved in 2 mL ethyl acetate and then Pd / C (10%) (18 mg) was added. The reaction mixture was hydrogenated for 16 hours. The reaction mixture was filtered and the filtrate was concentrated. The product was isolated by preparative TLC using ethyl acetate as solvent (68 mg). ¹ H NMR (CDCl 3): 1.75 (m, 1H); 1.91 (m, 2 H); 1.95 (m, 1H); 2.20 (m, 1H); 2.37 (m, 1H); 3.58 (m, 1H); 3.66 (m, 2 H); 3.81 (s, 2 H); 3.85 (m, 1H); 3.87 (s, 6H); 5.00 (t, 1H); 5.35 (dd, 1H); 5.41 (bs, 2H); 6.53 (s, 2 H); 8.39 (s, 1H).

(h) Preparation of 2- (4-fluorophenyl) -5- (3-hydroxyureidylpropoxy) tetrahydrofuran (compounds 14,15)

Compounds 14 and 15 were prepared from 135 and 136 using a procedure similar to Example 3 (g), substituting Compound 134 for Compounds 135 and 136. 1 H NMR (CDCl 3): 14 (trans): 1.72 (m, 1H) ; 1.93 (m, 3H); 2.20 (m, 1H); 2.38 (m, 1H); 3.58 (m, 1H); 3.67 (m, 2 H); 3.85 (m, 1H); 5.05 (t, 1H); 5.33 (dd, 1H); 5.48 (bs, 2H); 7.00 (t. 2H); 7.28 (m, 2 H); 8.48 (bs, 1H). 15 (cis): 1.92 (m, 2H); 2.01 (m, 1H); 2.10 (m, 2H); 2.26 (m, 1H); 3.53 (m, 1H); 3.64 (m, 2 H); 3.87 (m, 1H); 4.98 (m, 1H); 5.20 (dd, 1H); 5.43 (bs, 2 H); 7.01 (m, 2 H); 7.31 (m, 2 H); 8.43 (bs, 1H).

Example 4 Preparation of trans-2- {3- (N-hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (207)

The scheme for the synthesis of compound 207 is illustrated in Scheme 8.

• · a a and a and a and and and and· • a a and ·· and * and aa and a and and and aa and and and a and and and and and • · aa • a aa a and a aa

205

Scheme 8 l>,. *? <! · T οβ '* γ- «:

oi »»; to »ro ¹ oo«; c <'«}»;

G I »Π o co o * m:» »<

»; f · ♦;

*: »R at ♦: (*« <:

4

I * '' c i i i

AND

1 '* .i · ♦ tr.

ii _t :

(a) Preparation of 2- (t-butyldimethylsilyloxy) -5- (4-fluorophenyl) tetrahydrofuran (compound 202)

2-Hydroxy-5- (4-fluorophenyl) tetrahydrofuran (550 mg, 3.0 mmol), t-butyldimethylsilyl chloride (498 mg, 3.3 mmol) and imidazole (450 mg, 6.6 mmol) were dissolved in 2 mL dry DMF. This solution was stirred under dry argon overnight, poured into 200 mL water and extracted with 2: 1 ethyl acetate: hexane (3 x 100 mL). The combined organic extracts were washed with water (4 x 200 mL) and brine (100 mL), dried over sodium sulfate and evaporated to give 830 mg (93%) of 2- (t-butyl-dimethylsilyloxy) -5- (4-methylsilyloxy) -5- (4-ol). -fluorophenyl) tetrahydrofuran (202, mixture of cis and trans isomers) as a colorless oil that did not need further purification: ¹ H NMR (CDCl ₃ ): δ 7.40 - 7.50

Ί · (2H, m, minor izomerj, 7.25 7.35 (2HJ m, major isomer), 7.00 - 7.10 (2H, ia jakhlavní ták minor isomer), 5.71 ^-? 5.75 (1H, m, major isomer) * 5.59-5.62 (1H, m, minor isomer), 5.12-5.20 (IH, m, major isomer), 4.90-4.98 (IH, m, minor isomer), 2.40-2.55 (1H, m, both major and minor isomer); 2.05-2.17 (IH, m, both ^T 'major and minor isomer) , 1.87 - 2.00 (IH, m, both major and minor isomers), 1.67 1.70 (1H, m, both major and minor isomers), (0.92 (s, 9H 'both major and minor) isomer), 0.16 (s, 6H, both major and minor isomers) ^J '(b) ^S ' Preparation of trans-2- (3-Tetrahydropyranyloxy-1-butynyl) -5- (4-fluorophenyl) ) tetrahydrofuran (compound 204) ' ^A ' 2- (t-Butyldimethylsilyloxy) -5- (4-fluorophenyl) tetrahydrofuran (202, 593 mg, 2.0 mmol) was stirred in 10 µl of dry methylene chloride (degassed with argon before use) The solution was cooled to -70 ° C. Trimethylsilyl bromide (290 μΐ, 2.2 m mol) was added dropwise with stirring at the same temperature in an argon atmosphere. Stirring was continued for another 1.5 h to give 2-bromo-5- (4'-fluorophenyl) tetrahydrofuran (203) which was not isolated but was subsequently used without further purification (see below). ^! '= ..►. · ¹ *

In a separate flask, 3-tetrahydropyranyloxy-1-butyne (370 mg, 2.4 mmol) was dissolved in dry THF (5 mL). The solution was cooled to -60 ° C and n-butyllithium (1.0 mL, 2.4 mmol) was added dropwise with stirring at the same temperature under argon. Stirring continued another. The resulting solution was aspirated into the syringe and added after aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa2 - bromotetrahydrofuran (prepared above) at -70 ° C. Stirring was continued at -78 ° C for a further 0.5 hours. The reaction vessel was stored in a freezer (-78 ° C) overnight (although TLC showed no changes). The reaction mixture was poured into 2 M ammonium chloride solution (50 mL) and extracted with methylene chloride (3 x 50 mL). The solution was dried over sodium sulfate and the solvent was removed in vacuo. The residue was purified by flash column chromatography (eluent, 10% ethyl acetate in hexane) to give two products. Proton NMR analysis showed that the less polar component is trans-2- (3-tetrahydropyranyloxy) -1-butynyl) -5 (4-fluorophenyl) tetrahydrofuran (204.280mg, 45%) and the more polar component is a mixture of more than one substance . This mixture was discarded. 1 H NMR (CDCl 3): δ 7.27-7.30 (2H, m); 7.01 (2H, t,> 8.7 Hz), 5.09 (1H, t,> 7.1 Hz); 4.91 - 4.95 (2H, m); 4.57 - 4.64 (1H, m); 3.78 - 3.90 (1H, m); 3.50-3.60 (1H, m); 2.30 - 2.50 (2H, m); 2.05 - 2.17 (1H, m); 1.70 - 1.90 (3H, m); 1.50 - 1.65 (4H, m); 1.48 (3H, d,> 6.6 Hz).

(c) Preparation of trans-2- (3-hydroxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (compound 205) trans-2- (3-Tetrahydropyranyloxy) -1-butynyl) -5- (4- fluorophenyl) tetrahydrofuran (204, 280 mg, 0.9 mmol) was dissolved in methanol (15 mL). To this solution was added 50 mg of p-toluenesulfonic acid and the resulting solution was stirred for 45 minutes. Saturated sodium bicarbonate solution (10 mL) was added. After stirring for 5 minutes, the solution was added to 10 mL of water, diluted with 15 mL of brine, and extracted with methylene chloride (3 χ 30 mL). The combined organic phases were dried over sodium sulfate and the solvent was removed by rotary evaporation to give 212 mg (100%) of trans-2- (3-hydroxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (205). ^! 1 H NMR (CDCl 3): δ 7.29 (2H, dd,> 8.7 and 5.2 Hz); 7.01 (2H, t,> 8.7 Hz); 5.09 (1H, t,> 7.4 Hz); 4.92 (1H, t; J = 7.4 Hz); 4.59 (1H, q,> 6.6 Hz); 2.30'-2.50 (2H, m); 2.05 (t, 2 ', 1', 2,00, 1H, br s); 1.75 - 1.88 (1H, m); 1.47 (3H, d,> 6.6 Hz).

(c) Preparation of trans-2- {3- (N-phenoxycarbonyloxy-N-phenoxycarbonylamino) -1-butynyl} -5- (4-fluorophenyl) -tetrahydrofuran (compound 206):

trans-2- (3-Tetrahydropyranyloxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (205.210 mg, 0.89 mmol), triphenylphosphine (288 mg, 1.1 mmol) and N, O45 » • · • •......

Bis ((phenoxycarbonyl) hydroxylamine (283 mg, 1.1 mmol)) was dissolved in dry THF (5 mL) and cooled to 0 ° C under dry argon and dropwise. Diisopropyl azadicarboxylate (216 mL, 1.1 pmol) was added, stirring was continued for 1 hour at the same temperature, the solvent was evaporated and the residue was purified by flash column chromatography (eluent, 30% ethyl acetate in hexane) to give 250 mg (57%). %) trans-2- {3- (N-phenoxycarbonyloxy-N-phenoxycarbonylamino) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (206). 1 H NMR (CDCl 3): δ 7.15 - 7.45 (12H, m); 7.02 (2H, t, J = 8.6 Hz); 5.32 (1H, q J = 7.0 Hz); 5.07 (1H, t, J = 6.8) 4.96 (1H, t, J = 5.7 Hz), 2.25-2.50 (2H, m); 2.05-2.20 (IH, m); 1.70- 1.85 (1H, m); 1.66 (3H, d, J = 7.0 Hz).

(e) Preparation of trans-2- {3- (N-hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (compound 207) trans-2- {3- (N-phenoxycarbonyloxy-N-phenoxycarbonyl- amino) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (206.200 mg, 0.41 mmol) was dissolved in a high pressure tube in methylene chloride. The solvent was evaporated under a stream of argon and the residue was cooled to -78 ° C. Ammonia (8 mL) was condensed in this tube and 4 mL of t-butanol was added. The tube was sealed and allowed to warm slowly to room temperature over 18 hours. The pressure was released very slowly and the tube was left open for 1 hour. The residue was transferred to a flask and evaporated twice on a rotary evaporator with toluene added. The residue was purified by flash column chromatography (eluent, 3% methanol in ethyl acetate) and further purified by preparative TLC (solvent, 5% methanol in methylene chloride) to give 93 mg (78%) of trans-2- {3- (N-hydroxyuridyl) -1. -butinyl} -5- (4-fluorophenyl) tetrahydrofuran (207). IR (film) 3481, 3269, 2985, 2877, 2249, 1662, 1670, 1510, 1444, 1224, 1172, 1037 cm -1 ^! ; ^l HNMR

(1H, br s), 5.00-5.20 (2H, m); 4.80 - 5.00 (1H, m); 2.20 - 2.50 (2H, m); 1.70 - 2.20 (1H, m); 1.70 - 1.90 (1H, m); 1.37 (3H, dd, J = 6.9, 1.9 Hz).

Example 5 Preparation of S, S, S- and S, S, R-Isomers of trans-2- {3- (N-hydroxyureidyl) -butyl} -5- (4-fluorophenyl) -tetrahydrofuran (compounds 216 and 217)

• «· • · · • fe · fe · • fe • fe • • • fe · • fe ·· · * fe fe · • fefe • t · • • »» fe • • fe «· • fe fe * · • · • ·

One method for preparing the S, S, R- and S, S, S-isomers of trans-2- {3- (N-hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran is illustrated in Scheme 9.

208

3. PPTS

and.

OTBS

Diagram 9 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99 99

OCOOPH

PPhj, DiAD

219

OCOOPh ^, ΓΝ (S) / CH, O (R)

OPh

; 4N ^{N NH} (S) (S) / CH, o

220 (R)

Scheme 9 (continued) * * * * * * * * * * * * * * * * * 99 (a) Preparation of methyl 3- (4-fluorobenzoyl) propionate (compound 209)

To a solution of 3- (4-flurobenzoyl) propionic acid (1.98 g, 10.0 mmol) in methanol (25 mL) was added 0.5 mL of concentrated sulfuric acid. The resulting solution was stirred at ambient temperature under argon for 2 hours. The reaction mixture was neutralized with saturated sodium bicarbonate, methanol was removed on a rotary evaporator, and the residue was dissolved in 50 nml of ethyl acetate. The resulting solution was washed with saturated sodium bicarbonate (3 × 50 mL) and brine (50 mL), dried over sodium sulfate, and the solvent was removed in vacuo to give methyl 3- (4-fluorobenzoyl) propionate (2 g, 94%). IR (film) 3448, 3111, 3076, 3003, 3958, 1734, 1678, 1601, 1428, 1300, 1240, 1155, 1099 cm ^-1 ; 1 H NMR (CDCl ₃ ) δ 7.97 (2H, dd, J = 9.0, 5.5 Hz); 7.10 (2H, t, &gt; 8.9 Hz); 3.67 (3 H, s); 3.25 (2H, t,> 6.6 Hz); 2.73 (2H, t,> 6.6 Hz); ¹³ CNMR (CDCl3) δ 196,50; 173.34; 167.54; 164.17; 132.98; 130.77; 115.91; 115.62; 51.91; 33.31; 28,00.

(b) Preparation of (S) -5- (4-fluorophenyl) -y-butyrolactone (Compound 210)

A solution of methyl 3- (4-fluorobenzoyl) propionate (209, 780 mg, 3.67 mmol) in dry THF (2 mL) was added dropwise to a pre-cooled (0 ° C) solution of (-) - DIP chloride (2.02) g (6.25 mmol) in THF (2 mL) with stirring under a dry argon atmosphere. The starting solution was stirred at the same temperature for 2 hours and allowed to stand at 0-5 ° C overnight. The temperature was maintained at 0 ° C and water (2 mL) was added dropwise with stirring, followed by methanol (5 mL) and 5 M NaOH solution (5 mL). The reaction mixture was stirred at ambient temperature for 1.5 hours, cooled, and then 15 mL of saturated bicarbonate solution was added. The resulting mixture was washed with ether (3 x 50 mL) and acidified with 6N HCl. The acidified mixture was extracted with toluene (3 x 50 mL). The combined toluene extracts were washed with water and brine (50 mL), dried over sodium sulfate and the solvent was removed in vacuo. The residue was re-suspended in 50 mL of toluene and PPTS (10 mg) was added. The resulting solution was refluxed under a Dean-Stark trap (the first 15 mL of distillate was withdrawn) for 2 hours. The solution was cooled, washed with saturated bicarbonate solution (2 × 50 mL), dried over sodium sulfate, and the solvent was removed in vacuo to give 620 mg (94%) of (S) -5- (4-fluorophenyl) -y-butyrolactone. @ 1 H NMR (CDCl3) .delta

In.... I

999 I »♦ 9 * ··· · 9

9 9 9 9 9 9

999 99 99 999 ·· «·

7.33 (2H, dd,> 8.8, 5.3 Hz); 7.09 (2H, t,> 8.7 Hz); 5.50 (1H, dd,> 8.4, 5.9 Hz); 2.64 · 1 »

2.71 (3 H, m); 2.17-2.22 (1H, m).

(c) Preparation of (5S) -2-hydroxy-5-hydroxy-5- (4-fluorophenyl) tetrahydrofuran (compound 211) ((S) -5- (4-fluorophenyl) -y-butyrolactone (210.620 mg, 3, 44 mmol) was azeotroped (hexane) and dissolved in dry methylene chloride (25 mL), cooled to 78 ° C and DIBALH (3.5 mL of 1.5 solution in toluene, 5 mL) was added dropwise with stirring under argon. 25 mL of saturated b

sodium potassium tartrate solution. The cooling bath was removed and stirring continued for a further 2 hours. The reaction mixture was cooled with methylene chloride (25 mL). The organic layer was removed, washed with water (2 x 50 mL) and brine (50 mL), dried over sodium sulfate and the solvent was removed in vacuo to give 2-hydroxy-5- (4-fluorophenyl) tetrahydrofuran (620 mg, 100%). ^! 1 H NMR (CDCl ₃ ) δ 7.30-7.41 (m, 2H); 7.04 (m, 2 H); 5.63 - 5.78 (m, 1H); 5.00 - 5.22 (m, 1H); 2.48 (m, 1H); 2.20 - 2.32 (m,

IH); 1.95-2.10 (m, 1H); 1.79 (m, 1H).

(d) Preparation of (5S) -2- (t-butyldeimethylsilyloxy) -5- (4-fluorophenyl) tetrahydrofuran (compound 212) (5S) -2-hydroxy-5- (4-fluorophenyl) tetrahydrofuran (211.620 mg, 3, 5 mmol), t-butyldimethylsilyl chloride (700 mg, 5.25 mmol) and imidazole (595 mg, 8.75 mmol) were dissolved in 2 mL dry DMF, the resulting solution was stirred under argon overnight, poured into 200 mL water and extracted 2: 1 ethyl acetate: hexane (3 x 100 mL).

* The combined organic extracts were washed with water (4 x 200 mL) and brine (100 mL), dried over sodium sulfate and the solvent removed in vacuo to give 1 g (96 b ^1st%) of (5S) -2- (t-butyIdimethylsilyloxy ) -5- (4-fluorophenyl) tetrahydrofuran (212, a mixture of cis and _{Λ K.} I and ^A ..... trans trans isomers) as a colorless oil that did not need further purification. 1 H NMR - (CDCl 3) δ 7.40-7.50 (2H, m, minor isomer); 7.25 - 7.35 (2H, m, major isomer);

7.00-7.10 (2H, m, both major and minor isomer); 5.71 - 5.75 (1H, m, major isomer); 5.59 - 5.62 (1H, m, minor isomer); 5.12 - 5.20 (1H, m, major isomer);

4.90 - 4.98 (1H, m, minor isomer); 2.40 - 2.55 (1H, m, both major and minor isomer); 2.05 - 2.17 (1H, m, both major and minor isomer); 1,87 50 • ·· ·· · · «φ *» · · · · · · φ · · · · φ · φ ·

Φ Φ φ Φ »φ Φ φ

1 «Φ · ΦΦΙ φφ

2.00 (1H, m, both major and minor isomer); 1.67-1.70 (1H, m, both major and minor isomer); 0.92 (s, 9H, major and minor isomers); 0.16 (s, 6H, both major and minor isomer).

(e) Preparation of (2S, 5S) -trans-2- (3- t -butyldimethylsilyloxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (Compound 213) (5S) -2- (t-butyldimethylsilyloxy) -5- (4-Fluorophenyl) tetrahydrofuran (212, 1 g, 3.4 mmol) was dissolved in 10 mL dry methylene chloride (degassed by bubbling argon before use). This solution was cooled to -70 ° C and trimethylsilyl bromide (550 μΐ, 4.1 mmol) was added dropwise with stirring at the same temperature. Stirring was continued for another 1.5 hours to give (5S) -2-bromo-5- (4-fluorophenyl) tetrahydrofuran, which was used without further isolation (see below). In a separate flask, 3-t-butyldimethylsilyloxy-1-butine (840 mg, 4.5 mmol) was dissolved in dry THF (10 mL). The solution was cooled to -60 ° C and n-butyllithium (1.8 mL of a 2.5 M solution in hexane, 4.5 mmol) was added under a dry argon atmosphere while stirring at the same temperature. Stirring was continued for a further 0.5 hours. The resulting solution was added dropwise, rod-wise, to a stirred solution of 2-bromotetrahydrofuran (produced above) at -70 ° C. Stirring was continued at -78 ° C for an additional 1.5 hours. The reaction flask was left in the freezer (-78 ° C) overnight (although TLC showed no changes). The reaction mixture was poured into 2 M ammonium chloride solution (100 mL) and extracted with methylene chloride (3 x 75 mL). The solution was dried over sodium sulfate and the solvent was removed in vacuo. The residue was purified by flash column chromatography (eluent, 10% ethyl acetate in hexane) to give two components. Proton NMR analysis indicated that the less polar component was (2S, 5S) -trans-2- (3- t -butyldimethylsilyloxy-1-butyne) -5- (4-fluorophenyl) tetrahydrofuran (213.765 mg, 65%). 1 H NMR (CDCl 3) δ 7.29 (2H, m); 7.01 (2H, t, J = 8.7Hz); 5.09 (1H, t, J = 7.1 Hz); 4.91 - 4.97 (2H, m); 4.55 62 ^ ¹ * (IH, m); 2.26-2.50 (2H, m); 2.05 - 2.17 (1H, m); 1.38 (3H, d, J = 6.6Hz); 0.90 (9H, s); 0.12 (6 H, s). The more polar component was assigned to (2R, 5S) -cis-2- (3- t -butyl dimethylsilyloxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (214, 190 mg, 16%).

(f) Preparation of (2S, 5S) -trans-2- (3-hydroxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (Compound 215) 99

999 «9

9 * 9 9 · * 99 * 9 * 99 9 9 9 9 * 9 9 9 99 • 9 9 999 9 9

9,999-9,999- (25,55) -trans-2- (3- t -butyldimethylsilyloxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (213.765 mg, 2.2 mmol) was dissolved in 20 mL of Ti IF.Roztok was cooled to 0 C and ^IJ thereto was added TBAF (6.6 mL of 1M solution in THF). The resulting solution was stirred at 0 ° C for 2 hours and the solvent was removed in vacuo. The residue was dissolved in ethyl acetate (100 mL), washed with water (3 x 100 mL, 5 mL of brine added each time) followed by brine (50 mL), dried over sodium sulfate and the solvent removed in vacuo to give 500 mg (97 mL). %) (2S, 5S) -trans-2- (3-hydroxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (215). 1 H NMR (CDCl 3) δ 7.29 (2H, dd, J = 8.7, 5.2 Hz); 7.01 (2H, t,> 8.7 Hz); 5.09 (1H, t,> 7.4 Hz); 4.92 (1H, t,> 7.4 Hz); 4.59 (1H, q. &Gt; 6.6 Hz); 2.30 - 2.50 (2H, m); 2.05-2.15 (1H, m); 1.75 - 1.88 (1H, m); 1.72 (1H, br s); 1.47 (31 d, > 6.6 Hz).

(25.55) -trans-2- (3-Hydroxy-1-butynyl) -5- (4-fluoro-phenyl) -tetrahydro-furan (215. 500 mg, 2.13 mmol), (R) -α-methoxy-phenylacetic acid (1, 06 g, 6.4 mmol) and DMAP (86 mg, 0.7 mmol) were dissolved in dry methylene chloride (3 mL). DCC (1.5 g, 7.24 mmol) was added and the resulting solution was stirred at ambient temperature, under dry argon, for 3 hours (many white precipitates within minutes). The solid was filtered off and the filtrate was concentrated in vacuo. The residue was purified by flash column chromatography (eluent, 8% ethyl acetate in hexane) to give two diastereomeric esters. The less polar form was assigned (2S, 5S) -trans-2- {3- (S) -hydroxy-1-butynyl> 5 (4-thiophenyl) tetrahydrofuran (216, 250 mg, 30%,> 95% of 'H'). 1 H NMR (CDCl 3) δ 7.25-7.50 (7H, m); 7.02 (2H, t,> 8.5 Hz); 5.52-5.60 (1H, rn); 06 (1H, t, 1-6.8 liz); 4.88-4.94 (1H, m); 4.78 (1H, s); 3.43 (311, s), 2.25-2.47 (211, m); 2.00 2.13 (111, m); 1.75-1.88 (1H, m); 1.37 (3H, d,> 6.7 Hz). (2S, 5S) -trans-2- {3- (R) -hydroxy-1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (217, 230 mg. 29%, 72% by 1 H NMR). ¹ H NMR (CDCl ₃ ) δ 7.22-7.50 (7H, m); 7.01 (211, t, 1-8.7 Hz); 5.50 - 5.60 (1H, m); .98 (1H, t,> 7.2 Hz); 4.79-4.85 (1H, 1H); 4.79 (1H, s); 3.44 (3H, s); 2.20-2.40 (2H, s); 1.88-1.98 (1H, m); 1.72-1.80 (1H, m); 1.51 (311 d, > 6.7 liz). stirring in 10 mL of 1 M KOH solution at cthanolového ^50U C followed by usual workup) of these two esters to provide the corresponding alcohols (2S, 5S) -trans-2- {3- (S) -hydroxy-l -butinyl) -5- (452);

Aaa fluorophenyl) tetrahydrofuran (218, 150 mg, 98%) and its diastereomer (2S, 5S) -trans-2- {3 ( R) -hydroxy-1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (221, 50 mg, 100%). The 1 H NMR spectra of the two alcohols were identical to the spectrum measured for 218.

(g) Preparation of (2S, 5S) - (trans-2- {3- (R) - (N-Phenoxycarbonyloxy-N-phenoxycarbonylamino) -1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (Compound 219) (2S, 5S) -trans-2- (3- (S) -hydroxy-1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (218, 150 mg, 0.64 mmol), triphenylphosphine (200 mg, 0.77 mmol) and N, Obis (phenoxycarbonyl) hydroxylamine (200 mg, 0.77 mmol) were dissolved in dry TI IF (3 mL), cooled to 0 ° C, and propylazodicarboxylate (142 µL, Stirring was continued for 1 h at the same temperature The solvent was removed by rotary evaporation and the residue was purified by flash column chromatography (eluent, 30% ethyl acetate in hexane) to give 225 mg (72%) (2S, 5S). ) -trans-2- {3- (R) - (N- phenoxycarbonyloxy Nfenoxykarbonylamino) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran ^(219).! l 1 H NMR (CDCl) O 7.15 7.45 (12H, 1H), 7.02 (211, t, J = 8.6 Hz), 5.32 (1H, q, J = 7.0 Hz), 5.07 (1H, t .1-6.8 liz) 4.9 Δ (1H, t, J = 5.7 Hz); 2.25-2.50 (211. m); 2.05 - 2.20 (1H, m); 1.70 - 1.85 (1H, ni); 1.66 (3H, d, J = 7.0 Hz).

(h) Preparation of (2S, 5S) -trans-2- (3- (S) - (N-Phenoxycarbonyloxy-N'enoxycarbonylamino) -1-butynyl) -5- (4-fluorophenyl) tetrahydrofuran (Compound 222)

220 mg (70%) of the starting (2S, 5S) trans-2- {3- (R) -hydroxy-1-butynyl-5- (4- (1-fluorophenyl) tetrahydrofuran) was prepared according to the same procedure as for the preparation of compound 218. (2S, 5S) -trans-2- {3- (S) - (N-phenoxycarbonyloxy-N-phenoxycarbonylamino) -1butyl} -5- (4-fluorophenyl) tetrafluorofuran (222) .111 NMR spectra were the same as spectra for compound 219.

(i) Preparation of (2S, 5S) -trans-2- {3- (R) - (N-hydroxyurcidyl) -1-butynyl} -5- (4-fluorophenyl) -tetrahydrofuran (CMI-947) (Compound 220) ι * 9 * a 9 99 * a · a 9 a «a» aaa aaa * aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa (2S, 5S) “trans-2 - {3- (R) - (N-Phenoxycarbonyloxy-N-phenoxycarbonylamino) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (219.225 mg) was dissolved in a high pressure tube as a solution in methylene chloride. The tube was cooled to -78 [deg.] C. 10 ml of ammonia were liquefied in this tube and 2 ml of t-butanol was added, the tube was sealed and allowed to warm slowly to room temperature, then allowed to stir at room temperature for 18 hours. The pressure was then released very slowly and the tube was left open for 1 hour, the residue was transferred to a flask and concentrated twice in vacuo with toluene, and the residue was purified by preparative TLC (eluent, 5% methanol in methanol). thylene chloride) afforded 120 mg (90%) of (2S, 5S) -trans-2- {3- (R) - (N-hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (CMI-947,220). IR (film) 3209, 2985, 2874, 1653, 1510, 1449, 1326, 1224, 1117, 1037 cm ^-1 ; ^! 1 H NMR (CD ₃ OD) δ 7.34 (2H, dd, J = 8.7.5.4 Hz); 7.04 (2H, t, J = 8.8Hz); 5.00 - 5.10 (2H, m); 4.85 - 4.95 (1H, m); 2.25-2.50 (2H, m); 2.00-2.15 (1H, m); 1.78 - 1.85 (1H, m); 1.38 (3H, d, J = 7.0Hz).

(j) Preparation of (2S, 5S) -trans-2- {3- (S) - (N) -hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (CNI-948) (compound 223)

The same procedure as for the preparation of compound 219 yielded 110 mg of (2S, 5S) -trans-2- {3- (S) (N-phenoxycarbonyloxy-N-phenoxycarbonylamino) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran. (83%) (2S, 5S) -trans-2- {3- (S) - (N-hydroxyuridyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (CMI-948, 223). IR (film) 3200, 2985, 2881, 1643, 1510, 1442, 1222, 1035 cm -1; ¹ H NMR (CD ₃ OD) δ 7.34 (2H, dd, J = 8.7, 5.5 Hz); 7.04 (2H, t, J = 8.9Hz); 5.00 - 5.10 (2H, m); 4.85 - 4.95 (1H, m);

2.25-2.50 (2H, m); 2.00-2.15 (1H, m); 1.70 - 1.85 (1H, m); 1.38 (3H, d, J = 7.0Hz).

___, l d ... "-d * - at ......: *> - ·> -« »- ·:> -

Example 6 Preparation of the R, R, S- and R, R, R-Isomers of trans-2- {3- (N-hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran (compounds 234 and 236)

One method of preparing the S, S, R- and S, S, S-isomers of trans-2- {3- (N-hydroxyureidyl) -1-butynyl} -5- (4-fluorophenyl) tetrahydrofuran is illustrated in Scheme 10 below. .

·· «* ·

(.

4 4 4 4 4 5

444 4 44 4 · 4 · 4 4 · · · · 4 4 4

444 44

1.TMS-BT

I (ft) (A)

227

227

TBAF

H t (ft) (ft)

228

Scheme 10

OT8S

(ft) (S) _0H DCC. DMAP, (R) -α-methoxyphenylacetic acid

OTB $ I ·

229

OCOOPh θΗ ^ - ‘eooph ^ (fl) (fl, / CH, .ppi ^ oix?

(Sí

231

Scheme 10 (continued) (a) Preparation of 4- (4-Fluorophenyl) -4-oxomethylbutanoate (Compound 209)

To a stirred solution of 3- (4-benzoyl) propionic acid (208) (5.0 g) in methanol (20 mL) was added a few drops of sulfuric acid. After stirring overnight (19 hours), the reaction was neutralized with saturated aqueous sodium bicarbonate and the methanol was removed under reduced pressure. The rest was>

dissolved in ethyl acetate (50 mL) and washed with saturated aqueous sodium bicarbonate solution (3 x 15 mL), water (2 x 15 mL) and brine (2 x 15 mL), dried (Na ₂ SO 4), filtered and concentrated to give a light crystalline solid (5, 3 g,

%). ¹ H NMR: 2.79 (t, 2H); 3.30 (t, 2H); 3.71 (s, 2H); 7.14 (t, 2H); 8.02 (m, 2 H).

(b) Preparation of R-4- (4-fluorophenyl) -Y-butyrolactone (Compound 224)

To the cooled solution (0 ° C), stirred solution of (+) - DIP chloride (25 g, 77.9 mmol) in dry TFF (20 mL) was added ketoester 209 solution (10.07 g, 48.0 mmol) after argon. ) in dry THF (20 mL). The reaction was placed in a refrigerator (4 ° C) for 30 hours and then returned to an ice bath where it was stirred with the addition of water (10 mL), methanol (30 mL) and 10% NaOH _aq (60 mL). The ice bath was then removed. After all the ester had been hydrolyzed, a saturated aqueous solution of sodium bicarbonate (80 mL) was added. The aqueous layer was extracted with ether (2 x 100 mL) and then acidified to pH 2 and extracted with benzene (2 x 180 mL). To the combined benzene layers was added pyridinium p-toluene sulfonate (60 mg) and these were then heated under reflux using a Dean-Stark trap. After completion of the reaction, the benzene solution was washed with a saturated aqueous solution of sodium bicarbonate (150 mL), brine (2 x 50 mL), dried (Na ₂ SO 4), filtered and concentrated to give a white crystalline substance. precedent (7.92 g, 91%). 1 H NMR 2.10-2.25 (m, 1H); 2.68 (m, 3H); 5.50 (m, 1H); 7.08 (t, 2 H); 7.30 (m, 2 H).

.1 'M .... »<Ø> 1 II - L.. -AND.

(c) Preparation of cis- and trans-5R-5- (4-fluorophenyl) -2-hydroxytetrahydrofuran (compound 225) To a stirred solution of lactone 224 (7.25 g, 40.3 mmol) in dry toluene (50 mL) ), cooled in a dry ice-acetone bath, diisobutylaluminum hydride (1.5 M, in toluene) (1.5 eq., 40 mL) was added. When the reaction was complete, methanol (10 ml) was added slowly, then a saturated aqueous solution of sodium potassium tartrate (60 ml) and the water bath was removed. This solution was allowed to stand overnight (16 hours), the layers were separated and the aqueous fraction was extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with water (3 x 30 mL) and brine (3 x 30 mL), dried (Na 2 SO 4), filtered, and concentrated. The product was a colorless oil as a mixture of two stereoisomers.

(ca. 50/50) (6.32 g, 86%), 1 H NMR 1.7 (m, 1H); 1.9-2.3 (m, 2H); 2.42 (m, 1H); 3.60 (bs, 0.5H); 3.72 (bs, 0.5H); 4.98 (t, 0.5H); 5.20 (t, 0.5H); 5.60 (bs, 0.5H); 5.72 (m,

0.5H); 7.00 (t. 2H); 7.25 (m, 1H); 7.40 (m, 1H).

(d) Preparation of cis- and trans-5R-5- (4-fluorophenyl) -2-t-butyldimethylsilyloxy tetrahydrofuran (Compound 226)

To a stirred solution of lactol 225 (6.32 g, 34.7 mmol) in methylene chloride (140 mL) was added imidazole (1.1 leq, 38.2 mmol, 2.60 g) and TBDMS chloride (5.77 g). After stirring overnight, the reaction mixture was filtered and concentrated. The crude product was filtered through a silica plug to give a colorless oil that was a mixture of two diastereomers (ca. 2: 1) (9.61 g, 93%). 1 H NMR 0.14 (s, 6H); 0.92 (s, 9H); 1.7 (m, 1H); 1.9-2.2 (m, 2H); 2.4-2.5 (m, 1H); 4.9 (m, 0.33H); 5.16 (t, 0.66H); 5.59 (m, 0.33H); 5.71 (dd, 0.66H); 7.00 (m. 2H); 7.25 (m, 1.33H); 7.40 (m, 0.66H).

The presence of each configuration at the 5-position was determined for a sample of this compound using a chiral solvent reagent, [2,2,2-trifluoro-1- (9anthryl) ethanol, 2.2 mg substrate, 40 mg CSA]. These conditions showed that the compound 226 did not contain a detectable amount of 5S isomers.

The diastereomeric mixture of compound 226 (2.2 mg), in which position 5 was a racemic mixture, was treated with CSA (40 mg), from a 4.86-4.92 ppm (0.33H) multiplet to become two , 64-4.72 and 4.78-4.84 ppm. For the second diastereomer (same spectrum), a doublet of doublets of 5.66-5.70 ppm has become two sets of doublets of 5.64-5.68 and 5.70-5.74 ppm. For the chiral reduced compound, a smaller multiplet (w / CSA) showed up at 4.62-4.70 ppm and a doublet of doublets showed up at 5.68-5.70 ppm. Evidence for the existence of other isomers was not observed. (E) Preparation of 2R, 5R-trans-5- (4-fluorophenyl) -2- (3- t -butyldimethylsilyloxy-1-butynyl) tetrahydrofuran (Compound 227).

4 4 4 * 4 4 4 4 4 4 4 44 4 4 44 44 44 44 44 44 4 4 44 4 4 * 44

To a solution of Compound 226 (500 mg, 1.69 mmol) in dry degassed methylene chloride (10 mL) cooled to -78 ° C was added TMS bromide (0.25 mL, 1.86 mmol). This solution was stirred for four hours. To another flask containing 3-t-butyldimethylsilyloxy-1-butine (0.31 g, 1.68 mmol) and THF (5 mL) was added n-butyllithium (1.6 M in hexane, 1.26 mL, 2.02 mmol). After 30 minutes the bar solution was transferred to the above prepared solution. After two hours, the reaction mixture was poured into 2M aqueous ammonium chloride solution (25 mL) and extracted into methylene chloride (3 x 25 mL), dried (Na 2 SO 4), filtered and concentrated. Flash chromatography (5% ethyl acetate in hexanes) gave the trans product as a clear oil (280 mg, 48%). 1 H NMR 0.17 (d, 6H); 0.91 (s, 9H); 1.42 (d, 3H); 1.8 (m, 1H); 2.25-2.50 (m, 2H); 4.58 (m, 1H); 4.91 (m, 1H); 5.09 (m, 1H); 7.0 (t. 2H); 7.30 (m,

2H).

(f) Preparation of 2R, 5R-trans-5- (4-fluorophenyl) -2- (3-hydroxyl-1-butynyl tetrahydrofuran) (Compound 228)

To a stirred solution of 227 (0.38 g, 1.1 mmol) in THF (5 mL) cooled in an ice bath was added tetrabutylammonium fluoride (0.86 g, 3.3 mmol). The ice bath was removed. After 30 minutes the solvent was removed and the products were separated by flash ehromaiography (25% ethyl acetate in hexanes). The product was a colorless oil (170 mg, 67%). ¹ H NMR 1.48 (d, 3H); 1.8 (m, 1H); 2.1 (m, 1H); 2.3-2.5 (m, 2H); 4.58 (m, 1H); 4.91 (t, 1H); 5.1 (t, 1H); 7.0 (t. 2H); 7.29 (m, 2 H).

The hydroxy function of 228 was esterified with R-methoxyphenylacetic acid (DCC, DMAP, _CH2 Cl2, 55% after chromatography) and the resulting diastereomers (229 + 230) were separated (flash chromatography) and were isolated as the R and S isomers at the carbinol carbon. The ester was removed by basic hydrolysis (KOH, 78%) to give carbinols 231 and 232. Absolute configurations were assigned based on the Mosher model.

(g) Preparation of 2R, 5R-trans-5- (4-fluorophenyl) -2- (3R-3-N, O-bisphenoxycarbonyl hydroxylamino-1-butynyl) tetrahydrofuran (Compound 233)

To the cooled (ice bath) solution of 2R, 5R-trans-5 - ((4-fluorophenyl) -2- (3S-3hydroxy-1-butynyl) tetrahydrofuran (231) (29 mg, 0.12 mmol), triphenylphosphine (39). mg, • · I * i i ·· ··

0.15 mmol) and N, O-bisphenoxycarbonylhydroxylamine (37 mg, 0.14 mmol) in THF (3 mL) were slowly added diisopropylazodicarboxylate (0.029 mL, 0.15 mL). The ice bath was removed and when the reaction was complete (a few minutes) the solvent was removed. The product was obtained by flash chromatography (15% ethyl acetate in hexanes) as a colorless oil (32 mg, 53%). 1 H NMR 1.65 (d, 3H); 1.8 (m, IH); 2.1 (m, 1H); 2.4 (m. 2H); 4.94 (m, 1H); 5.08 (m, 1H); 5.30 (m, 1H); 7.0 (t. 2H); 7.15-7.40 (m, 2H).

(h) Preparation of 2R, 5R-trans-5- (fluorophenyl) -2- (3R-3-N-hydroxyureidyl-1-butynyl) tetrahydrofuran (Compound 234)

Compound 233 (32 mg) was placed in a vial (-78 ° C) with screw cap and stir bar along with liquid ammonia (ca. 3 ml) and t-butanol (ca. 2 ml). The vessel was sealed and the cold bath removed. After stirring overnight at room temperature, the pressure was released and the solvent was removed. The product was triturated (25% ethyl acetate in hexanes) to give a white solid (14 mg, 74%). 1 H NMR 1.41 (d, 3H); 1.8 (m, 1H); 2.1 (m, 1H); 2.3-2.5 (m, 2H); 4.93 (t, 1H); 5.08 (t, 1H); 5.20 (m, 1H); 5.38 (bs, 1H); 7.0 (t. 2H); 7.29 (m, 2 H).

Electron Spray MS: M + 1 = 293

The synthesis of the RRS isomer (CMI-957) was carried out in the same manner from ester 230 as the RRR isomer (CMI-954) was prepared from ester 29.

Example 7 Preparation of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4-N-hydroxyureidyl) butynyl) tetrahydrofuran (compound 1, Figure 4) and 2S, 5R-trans-2- (4-fluorophenoxymethyl) -5- (4-N-hydroxyureidylbutyl) tetrahydrofuran (Compound 402, Figure 4)

Preparation of 4S- (4-fluorophenoxymethyl) -γ-butyrolactone (Compound 301, Figure 4)

To a stirred solution of (S) -γ-butyrolactone (1.0 g, 8.61 mmol), 4-fluorophenol (1.16 g, 10.35 mmol) and triphenylphosphine (2.49 g, 9.49 mmol) in 10 ml THF was added dropwise diisopropyl azadicarboxylate (1.87 µΐ, 9.46 mmol). After the addition was complete, the reaction mixture was stirred at 80 ° C for 16 hours. The solvent was then removed and the product was separated by flash column chromatography (silica, 2: 1 in 4: 4 in silica gel).

4 hexane / ethyl acetate) (1.38 g, 76.3%). 1 H NMR (CDCl ₃ ) 2.27 (m, 1H); 2.42 (m, IH); 2.60 (m, IH); 4.04 (m, IH); 4.15 (m, IH); 4.85 (m, 1H); 6.84 (m, 2 H); 6.98 (m, 2 H).

Preparation of 2S- (4-Fluorophenoxymethyl) -5-hydroxytetrahydrofuran (Compound 302, Figure 4).

To a stirred solution of lactone 301 (1.38 g, 27.22 mmol) in dry toluene (24 mL) at -78 ° C was added dropwise a 1.5 M toluene solution of DIBALH (6.76 mL, 10.13 mmol). . The reaction mixture was stirred at -78 ° C for 2 hours. The reaction mixture was cooled by addition of methanol (1.7 mL) at < -60 ° C. The mixture was warmed to -20 ° C, followed by the addition of a saturated aqueous solution of sodium potassium tartrate (10 mL), maintaining the temperature of the reaction mixture between -10 and 0 ° C. The reaction mixture was stirred at room temperature overnight and then the two phases were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, saturated NaCl solution and concentrated in vacuo to give an oil which was purified by flash column chromatography (silica, 1: 1 hexane / ethyl acetate (1.41 g, 101%). * HNMR (CDCl ₃ ) 1) 80 (m, 1H); 2.05 (m, 2H); 2.26 (m, 1H); 3.93 (m, 2H); 4.04 (m, 2H); 4.47 (m, 1H); 0.5H), 4.61 (m, 0.5H), 5.57 (m, 0.5H), 5.66 (m, 0.5H), 6.88 (m, 2H), 6.98 (m, 2H).

Preparation of 2S- (4-fluorophenoxymethyl-5- (t-butyldimethylsilyloxy) tetrahydrofuran (Compound 303, Figure 4)

To a stirred solution of lactol 302 (1.41 g, 6.65 mmol) in methylene chloride (25 mL) was added imidazole (498.1 mg, 7.32 mmol) and TBDMS chloride (1.1 g, 7.32 mmol). . The reaction mixture was stirred at room temperature overnight and then the reaction mixture was filtered and the filtrate was concentrated. The crude product was purified by flash column chromatography using 9: 1 hexane / ethyl acetate as solvent to give a colorless oil which was a mixture of two diastereomers (ca. 2: 1) (1.22 g, 56.2%). 1 H NMR (CDCl ₃ ) 0.11 (s, 6H); 0.90 (s, 9H); 1.80-2.10 (m, 3H); 2.22 (m, IH); 3.91 (m, 2 H); 4.38 (m, O, 33H); 4.50 (m, 0.67H); 5.52 (m, 0.33H); 5.59 (m, 0.67H); 6.86 (m, 2 H); 6.96 (m, 2 H).

Fr · fr fr fr fr »» *** *** *** *** *** *** *** *** *** *** *** ***

Preparation of 2S, 5S-trans-2- (4-fluorophenoxymethyl-5- (4-t-butyldimethylsilyloxy-1-butynyl) tetrahydrofuran (compound 304, Figure 4) and 2S, 5R-cis-2- (4-fluorophenoxymethyl) -5- (4- butyldimethylsilyloxy-1-butynyl) tetrahydrofuran t. (Compound 305, Figure 4)

To a stirred solution of 303 (720 mg, 2.21 mmol) in dry methylene chloride (10 mL) cooled to -78 ° C was added TMS bromide (349.8 μΐ, 2.65 mmol). The reaction mixture was stirred at -78 ° C for 4 hours. In a separate flask containing 4-t-butyl-dimethylsilyloxy-1-butine (812.8 mg, 4.42 mmol) and THF (10 mL) was added n-butyllithium (2.5 M in hexane, 2.65 mL, 6.63 mmol). After 30 minutes it was transferred to the solution prepared above. After two hours, the reaction was quenched by addition of saturated aqueous ammonium chloride solution, and the mixture was extracted with methylene chloride, dried over MgSO ₄ , filtered and concentrated. Flash column chromatography (silica, 95: 5 hexane / ethyl acetate) yielded two products, trans (304) (210 mg) and cis (305) (160 mg) and a mixture of the two (50 mg). The overall yield was 48.5%. 1 H NMR (CDCl ₃ ) 304: 0.10 (s, 6H); 0.91 (s, 9H); 1.87 (m, 1H); 2.01 (m, 1H); 2.22 (m, 2 H); 2.43 (t, 2 H); 3.72 (t, 2 H); 3.92 (d, 2 H); 4.47 (m, 1H); 4.73 (m, 1H); 6.86 (m, 2 H); 6.95 (t, 2 H). 305: 0.09 (s. 6H); 0.90 (s, 9H); 1.92 - 2.20 (m, 4H); 2.42 (m, 2 H); 3.70 (t, 2 H); 3.92 (m, 1H); 4.07 (m, 1H);

4.29 (m, 1H); 4.62 (m, 1H); 6.86 (m, 2 H); 6.96 (t, 2 H).

In the preparation of compounds 304 and 305, an oxygen protecting group, 4-t-butyldimethylsilyl, may be replaced by another as known to those skilled in the art.

»Ί

In order to determine the stereochemistry of this molecule, a different NOE experiment was performed for both 304 and 305. In the difference experiment 304, a multiplet at 4.73 ppm would be irradiated with a low-frequency rf pulse disconnect pulse and

*. U r -, j ·, 1, - 1 -, ,, H II - ^ »lh - H ...

the data was processed so that only the signal increase could be observed. This represents a positive NOE effect and suggests a close spatial relationship of these protons.

In this experiment, a NOE was found for a 2.22 ppm multiplet, which is a turanate proton, a 4.47 ppm multiplet was irradiated with a low-frequency pulse, and the data was processed so that only the signal increase was measured. The NOE effect has been found for • • • • • * * • • • * * · · · · · «♦ · t t t t t t t t multiplet with 2.22 ppm, which are protons of the furan ring. Another NOE was observed for 3.92 ppm protons, which are protons on methylene following this multiplet.

In the differential NOE experiment of Compound 305, a triplet at 4.62 ppm was irradiated with a low-frequency rf spin disconnect pulse and the data was processed so that only signal magnification was measured. This represents a positive NOE effect, suggesting a close spatial relationship of these protons. In this experiment, NOE was found at 4.29 ppm, the second methine furan proton. Additional NOE was observed for the multiplet at 2.17 ppm, which are furan protons. Another NOE was found for the triplet at 4.62 ppm, the second methine furan proton. Additional NOE was observed for protons at 3.92 and 4.07 ppm, which are protons on methylene following this multiplet. Additional NOE was observed for the multiplet at 2.11 ppm, which are furan protons.

Preparation of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4-hydroxy-1-butynyl tetrahydrofuran) (Compound 306, Figure 4)

To a stirred solution of 304 (210 mg, 0.54 mmol) in THF (1.4 mL) cooled in an ice bath was added tetrabutylammonium fluoride (420.3 mg, 1.61 mmol). The ice bath was removed and the reaction stirred at room temperature for 1 hour. The solvent was removed and the product was separated by flash column chromatography (silica, Lhexane / ethyl acetate) (124 mg, 83.2%). 1 H NMR (CDCl ₃ ) 1.88 (m, 1H); 2.02 (m, 1H); 2.25 (m, 2 H); 2.50 (m, 2 H); 3.72 (t, 2 H); 3.93 (d, 2 H); 4.48 (m, 1H); 4.76 (m, 1H); 6.84 (m, 2 H); 6.96 (t, 2 H).

Preparation of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4-N, obisphenoxycarbonylhydroxyamino-1-butynyl) tetrahydrofuran (Compound 307, Figure 4) u - *. .........

To a cooled solution (ice bath) of compound 306 (124.0 mg, 0.45 mmol), triphenylphosphine (128.9 mg, 0.49 mmol) and N, O-bisphenoxycarbonylhydroxylamine (147.3 mg, 0.54 mmol) in tHF (5 mL) was added diisopropoxylazodicarboxylate (94.1 μΐ, 0.48 mmol). The ice bath was removed and the reaction mixture was warmed to ambient temperature and stirred at this temperature for 30 minutes. The solvent was removed and the product was purified by flash column chromatography (silica, 4: 1 → 44).

5 · * * ·

444 ♦ · 3 · ·

44 4

4 * 44

4 4 4

4 44

44 4 4

4>

444 hexane / ethyl acetate (195 mg, 82.0%); ¹ H NMR (CDCl 3) 1.85 (m, 1H); 2.03 (m, 1H); 2.22 (m, 2 H); 2.75 (m, 2 H); 3.92 (d, 2 H); 4.05 (m. 2H); 4.47 (m, 1H); 4.76 (m, 1H);

6.84 (m, 2 H); 6.95 (m, 2 H); 7.26 (m, 5H); 7.41 (m, 5H).

Preparation of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4-N-hydroxyureidyl) -1-butynyl) tetrahydrofuran (Compound 401, Figure 4)

In a screw cap flask was placed NH 3 at -78 ° C (approximately 1-2 mL). Compound 307 (195.0 mg, 0.37 mmol), previously dissolved in 20 mL of methanol, was added to this cold liquid ammonia. The flask was sealed and the ice bath was removed. The reaction mixture was stirred at ambient temperature for 16 hours. The reaction mixture was recooled in a dry ice bath and pressure was released. The flask was opened and the solvent was removed. The product was isolated by flash column chromatography using ethyl acetate as solvent to give a solid (108 mg, 91.7%). NMR (CDCl 3) 1.84 (m, 1H); 2.01 (m, 1H); 2.22 (m, 2 H); 2.55 (t, 2 H); 3.75 (t, 2 H); 3.94 (m, 2 H); 4.48 (m, 1H); 4.74 (t, 1H); 5.25 (bs, 2 H); 6.86 (m, 2 H); 6.98 (m, 2 H).

Preparation of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4-N-hydroxylureidyl-1-butynyl) tetrahydrofuran (Compound 402, Figure 4)

Compound 1 (75 mg, 0.23 mmol) was dissolved in 2 mL of ethyl acetate and then Pd / C (10%) (15 mg) was added and the reaction mixture was hydrogenated under balloon pressure for 16 hours. The reaction mixture was filtered and the filtrate was concentrated. The product was isolated by flash column chromatography using ethyl acetate as solvent (70 mg, 92.2%). 1 H NMR (CDCl ₃ ) 1.50-1.70 (m, 8H); 2.10 (m, 2H); 3.58 (m, 2 H);

3.91 (m / 2H), 4.08 (m, 1H), 4.40 (m, 1H); 5.15 (bs, 2Η); ^, 87 ^ Γ2Η); * 6.97 (ζ2Η);

7.40 (bs, 1H).

II. Pharmaceutical compositions

Humans, horses, dogs, cattle, and other animals, especially mammals suffering from inflammatory diseases, particularly those that are mediated by PAF or 564. Lipoxygenase can be treated by administering effective amounts of one or more of the above-described compounds or pharmaceutically acceptable derivatives or salts thereof. , also in the form of pharmaceutically acceptable carriers or diluents, wherein the treatment reduces the formation of oxygen radicals. These active materials can be administered by any suitable route, for example, orally, parenterally, intravenously, skin, or in other material forms such as liquid, cream, jelly, solid or aerosol form.

Accordingly, as used herein, the term pharmaceutically acceptable salt refers to salts or complexes that retain the desired biological activity of the above compounds and exhibit minimal undesirable toxicological effects. Examples of such salts, although not the only ones, are (a) acid salts formed by reaction with inorganic acids (eg hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.) and salts formed by reaction with organic acids such as e.g. acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid and polygalacturonic acid; (b) basic salts formed from metal cations such as zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, nickel, cadmium, sodium, potassium etc. or with cations formed from ammonia, Ν, d-dibenzylethylenediamine, D -glucosamine or ethylenediamine; (c) combinations of a) and b) eg tin (II) zinc salt, etc. The compounds may also be administered as pharmaceutically acceptable quaternary salts known to those skilled in the art that specifically include ammonium salts of the formula -NR 3 ⁺ Z ', wherein R is alkyl or benzyl and Z is a counter ion, including chloride, bromide, iodide, alkyl-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate or carboxylate (such as benzoate, succinate, acetate, glycolate, malate, citrate, tartrate, ascorbate, benzoate, cinnamon, mandlan , benzyloate a., -. ', .jj -. r - <··''* * - ··· if - · · - Φ ** ··. ^ I4i Itai ml ->

diphenylacetate).

The active compound is contained in an acceptable carrier or diluent in an amount sufficient to provide the patient with a therapeutically effective amount that will not cause any serious toxic effects in the patient. A preferred dose of the active compound for all of the above compounds is in the range of 10 ng / kg to 300 mg / kg, preferably 0.1 to 100 mg / kg per day, more generally 0.5 to 25 mg. · »Fe fe fe fe» fe ef ef »» »» »» »» »» »» • • • • • • • • • • • • • • fe fefe per kilogram weight recipient. A preferred dose for cardiovascular indications ranges in the range of 10 ng / kg to 20 mg / kg. A typical dose is in the range of 0.01 3% weight / weight in a suitable carrier. The effective dosage range can be calculated based on the weight of the parent substance to be delivered. If both the derivative exhibits activity, the effective dosage can be estimated as above using the weight of the derivative or other means known to those skilled in the art.

The compound may conveniently be administered in any suitable dosage unit form, such as 1 - 3000 mg, preferably in the range of 5 - 500 mg of the active ingredient f.

per unit dosage form. An oral dose of 25-250 mg is usually preferred.

The active ingredient is preferably administered to achieve a peak plasma concentration of about 0.00001-30 mM, preferably 0.1-30 μΜ. This can be achieved, for example, by intravenous injection of a solution of the active ingredient, preferably in saline or aqueous medium, or by administering large pills of the active ingredient.

The concentration of the active ingredient in the dosage form depends on the absorption, distribution, inactivation and excretion ratios of the drug, as well as other factors known to those skilled in the art. It is also worth noting that the dosage values will also vary depending on the severity of the problems to be alleviated. It should also be understood that for any specific subject, dosage regimens should be promptly adjusted according to the individual needs and professional judgment of the person administering or administering the drug, and that the concentration ranges given herein are exemplary only and are not intended to limit the extent of use of the claimed drug. The active ingredient may be administered at once or may be divided into small doses which will then be administered at different time intervals.

Oral compositions typically include an inert diluent or an edible carrier. These may then be in the form of gelatin capsules or compressed into tablets. For oral therapeutic administration, the active ingredient may be combined with excipients and used in the form of tablets, lozenges or capsules. Pharmaceutically compatible binders and / or adjuvants can also be used as part of a dosage form.

• fefe fefe · fefe fe fe fefe · fefe · fe · fefe · 4 · fefe · fefe fefe fe · fe · fe · fe • fe

Tablets, pills, capsules, lozenges, etc. may contain any of the following ingredients or compounds of a similar nature: a binder such as microcrystalline cellulose, a bovine resin or gelatin, excipients such as starch or lactose, a dispersing component such as alginic acid, Primogel or corn starch, a lubricant such as magnesium stearate or sterots, glidants such as colloidal silica, a sweetening agent such as sucrose or saccharin, or a flavoring agent such as peppermint, methyl salicylate or orange flavor. When the dosage unit form is a capsule, it may contain, as additional material to the materials mentioned above, a liquid carrier or a fatty oil. Further, dosage unit forms may contain other materials which may modify the physical form of the dosage unit, e.g., sugar coating, shellac, or enteric agents.

The active compound or pharmaceutically acceptable salt or derivative may be administered as part of an elixir, suspension, syrup, wafer, chewing gum, etc. In addition to the active compound, the syrup may contain sucrose as a sweetening agent or certain preservatives, colorants, coloring agents and flavors.

The active compound or pharmaceutically acceptable derivatives or salts may be admixed with other active materials that do not reduce the desired effect or with materials that supplement the desired effect, such as antibiotics, antifungal agents, other anti-inflammatory agents, or antiviral compounds.

Solutions or suspensions for parenteral, dermal or fabric applications may contain the following components: sterile diluent such as water for injection, saline, fixed oils, polyethylene glycols or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparagens; antioxidants such as ascorbic acid, sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid. buffers such as citrates áčetátýj ^J "nebo'fosfáty'nebo agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral formulations may be contained in ampoules, disposable injections or multiple tubes made of glass or plastic.

When administered intravenously, the preferred carriers are saline or phosphate buffered saline (PBS), and &quot; and &quot; and &quot; &quot; a * «a a.

aa aaa aa aa aa aaa aaa a aa a aa aa

In one modification, the active compounds are formulated with carriers that protect the compound from rapid elimination from the body, such as release controlling agents including implants and delivery systems. Biodegradable, biocompatible polymers can also be used, e.g., ethylene vinyl acetate, polyanhydrides, polyglycolic acids, collagen, polyorthoesters, and polyactic acid. Methods for preparing such systems are apparent to those skilled in the art. The substances can also be obtained commercially from Alza Corp. (CA) Scios Nova (Baltimore, MD).

Liposomal suspensions may be pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, e.g. as described in U.S. Pat. Patent No. 4,522,811 (which is incorporated herein by reference). For example, the liposomal composition can be prepared by dissolving a suitable liquid (eg, fosafitidylethanolamine, stearoylphosphatidylcholine, arachadoylphosphatidylcholine, and cholesterol) in an inorganic solvent, which is then evaporated and a dry lipid film is obtained on the surface of the vessel. An aqueous solution of the active compound or its monophosphate, diphosphate and / or triphosphate is introduced into a container. The contents of the container are then vortexed manually to release the lipid material from the sides of the container and disperse the lipid aggregates, thereby forming a liposomal suspension.

III. Biological activity

There are a variety of biological assays that have been used to evaluate the ability of a compound to play the role of a PAF receptor antagonist, including the ability of the compound to bind to PAF receptors and the effect of the compound on different PAF mediated pathways. Any of these assays can be used to evaluate the ability of the compounds disclosed herein to play the role of PAF receptor antagonists.

For example, it is known that PAF induces hemoconceration and increases the permeability of microcirculation leading to a decrease in plasma volume. Acute circulatory collapse caused by PAF can be used as a basis for attempting to evaluate the ability of a compound to play a PAF antagonist role by analyzing the effect of the compound on PAF-induced reduced plasma volume in an animal model such as mice.

• ΦΦ · * »» · φ φ φ φ · · · · · · · · · · · · · · · · · ·

ΦΦ

ΦΦ

Endotoxemia causes the release of chemical mediators including icosanoids, PAF, and tumor necrosis factor (TNF), which stimulate a variety of physiological responses including influenza, hypotension, leukocytosis, and glucose and fat metabolism disorders. Endotoxemia can lead to severe shock and death. Mice induced endotoxin induction is an important animal model for evaluating the pharmacological effects of compounds on endotoxic shock.

The other two common assays used to evaluate the compound as a PAF receptor antagonist are in vitro platelet aggregation and hypotension in rats (Shen, et al., "Chemical and Biological Properties of PAF Antagonists and Biosynthetic Inhibitors", Factor Activating Platelets and Related Fat mediators, F, Snyder, Ed Plenum Press, New York, NY 153 (1987)).

A variety of biological experiments were used to evaluate the compound's ability to inhibit the 5-lipoxygenase enzyme. For example, the rat basophilic leukemia cell cytosol 5-lipoxygenase has been widely used in leukotriene biosynthesis studies. Compounds that inhibit 5-lipoxygenase reduce leukotriene levels.

Another biological procedure used to evaluate the compound's ability to inhibit the 5-lipoxygenase enzyme is based on the classical pharmacological model in which inflammation is induced by inhibition of LTB ₄ by the human blood-stimulated ionophore.

Example 5 Ability of a compound to bind to PAF receptors (a) Preparation of human platelet membranes. n- -WF www I p.

Human platelet membranes are prepared from platelet concentrates obtained from American Red Cross Blood Services (Dedham, MA). After several washes with plate wash solution (150 mM NaCl, 10 nM Tris and 2 mM EDTA, pH 7.5), the plate granules were resuspended in 5 mM MgCl 2, 10 mM Tris and 2 mM EDTA at pH 7.0. The cells were then rapidly frozen with liquid nitrogen and then slowly thawed at room temperature. Freezing and thawing were at least three times

4 · 4 · · · ·

443 4 4 · ·

4 · »• 4 44 4 ** • 4 44 * 4 4 4 •« 44 • 44 4 4 • 4 4 • 4 44 repeated. For further fractionation of the membrane fragments, the disrupted membrane suspension was overlaid on top of a discontinuous sucrose gradient of 0.25, 1.03 and 1.5 M, prepared in 10 mM, 10 mM Tris and 2 mM EDTA, pH 7.0 , and then centrifuged at 63,500 µg for 2 hours. Membrane fractions forming bands between 0.25 and 1.03 M (membrane A) and between 1.03 and 1.5 M (membrane B) were collected separately. Protein concentration of the membrane preparations was then determined by the Lowry method using bovine albumin serum (BSA) as a standard. The membranes are then separated into smaller fractions (4 ml each), stored at -80 ° C and thawed before use.

(b) Inhibition of [ ³ H] PAF binding

The ability of [ ³ H] PAF to bind to specific receptors on human platelet membranes was evaluated under optimal conditions at pH 7.0 in the presence of 10 mM MgCl 2. To 0.5 ml of the final solution containing 0.15 pmol (0.3 nM concentration) of [ ³ H] PAF is added membrane protein (100 µg) and a known amount of unlabeled PAF or PAF receptor antagonist in 10 mM MgCl 2, 10 mM Tris, and 0.25% at pH 7.0. After incubation for four hours at 0 ° C, bound and unbound PAF were separated using a Whatman GF / C glass fiber filter. Under these conditions, no degradation to the filter bound [ ³ H] PAF was detected. Non-specific binding is defined as total binding in the presence of excess unlabeled PAF (1 mM) where no further displacement of either unlabeled PAF or PAF analogs or PAF receptor antagonists was found. Specific binding is defined as the difference between total binding and non-specific binding.

To determine the relative potency of test substances, the binding of [ ³ H] PAF in the presence of inhibitors is expressed by percent inhibition by placing total binding in the absence of inhibitors as 0% inhibition and total binding in the presence of 1 mM unlabeled PAF as 100% . Percent inhibition by a compound can be calculated according to the following expression:

Inhibition * [ftl% inhibition = [(total binding - total binding in the presence of the compound)] ) / nonspecific binding] x 100%

The IC 50 is then calculated as the inhibitor concentration required to obtain 50% inhibition of □ specific binding of [H] PAF and is calculated using the non-linear regression computer program GraphPad Inplot, version 3.0 (GraphPad Software, San Diego, CA).

Example 6 Effect of Compound on PAF Induced Hemoconcentration (a) Animals

Female CD-I mice weighing 16-20 grams were obtained from Charles River Laboratory (Wilmington, MA). Tap water and laboratory rodent food were provided (5001, Purina Mills, St. Louis, MO). Mice were housed for an average of four days before being used.

(b) Hematocrystalline measurement

PAF (1-O-alkyl-2-acetyl-1H-glyceryl-3-phosphorylcholine, Sigma Chemical, Co.) was dissolved in 0.25% bovine albumin serum (BSA) in 0.9% NaCl solution.

With the exception of dose response studies, 10 µg (10 ml / kg) of PAF solution was injected into the tail vein. All test compounds were dissolved in 0.5 DMSO saline and injected intravenously at 3 mg / kg body weight 15 minutes prior to PAF treatment. Thirty to fifty μΐ of blood is collected by cutting the tail end into a heparinized microhaematocrystal tube (1.50 mm diameter) 15 minutes after PAF administration. All test compounds were administered intravenously at 3 mg / kg body weight 15 minutes prior to PAF (10 µg / kg, intravenous) or AA (0.5 mg / ear) into the mouse.

Example 7 Effect of Compounds on Cytosol 5-Lipoxygenase of Rat Basophil Leukemia Cells (a) Preparation of Enzyme

Washed rat RBL cells (4 x 10 8) were suspended in 20 ml of 50 M potassium phosphate buffer at pH 7.4 containing 10% ethylene glycol / 1 mm EDTA (buffer A). The cell suspension was sonicated at 20 kHz for 30 seconds and the resulting sonicate was centrifuged at 10,000 xg for 10 minutes, followed by fefe feefefefe fefe fefe fefe fefe fefe fefe fefe fefe fefe • feef feefefef feefefefefefefefefefefeed in a further centrifugation at 105,000 xg for 1 hour. The supernatant solution (cytosolic fraction) containing 5-lipoxygenase was stored at -70 ° C. Protein concentration was determined according to the Bradford Dye Reagent procedure with bovine albumin serum as standard.

(c) Enzyme Assay

For routine determination of 5-lipoxygenase, the mixture contained 50 mM potassium phosphate buffer pH 7.4.2 mM CaCl 2, 2 mM ATP, 25 mM ATP, 25 M arachidonic acid (0.1 Ci) and enzyme (50-100 mg protein) in a final volume of 200 ml. The reaction was carried out at 24 ° C for 3 minutes. The mixture was extracted with 0.2 mL of ice-cold ethyl ether: methanol 0.2 M citric acid (30: 4: 1). Extraction was performed by thin layer chromatography at -10 ° C in a petroleum ether: ethyl ether: acetic acid (15: 85: 0.1) solvent system. The silica gel zones corresponding to the authentic arachidonic acid and its metabolites were scraped into scintillation tubes for further counting. Enzyme activity was expressed as the amount of arachidonic acid oxidized for 3 minutes. Compounds 9, 11, 14 and 15 mentioned above show activity in this experiment.

Example 8 Inhibition of Soluble 5-lipoxygenase in RBL-1 Cell Extract

RBL-1 cells were grown to confluency (2 x 10 ⁶ / ml) in flasks as specified by the ATCC. Cells are harvested and washed twice in PBS without calcium and magnesium. Cells were suspended at 2 x 10 ⁷ / ml in 50 mM K 2 HPO 4 pH 7.4, 10% PEG8000, 1 mM EDTA, 1 mM PMSF, and then sonicated. The resulting mixture was centrifuged at 100,000 xg for 1 hour at 4 ° C and the supernatant (cytosol) was collected and stored in aliquots at -70 ° C.

The activity of 5-L0 in cytosol RBL-1 was determined as follows: 0.2 ml reaction mixture consisting of 5 mM CaCl ₂ , 2 mM ATP, 50 mM K 2 HPO 4 pH 7.4, different concentrations of test substance (from 10 ml DMSO soluble compound) ) and the concentration of cytosol RBL-1, which converted 50% of the ¹⁴ C arachidonic acid substrate into oxidized products (experimentally determined for each cytosolic concentration), was &quot; t &quot; · ** ** ** ** ** ** ** ** ** ** ** ** ** ** · · · · · · · · · ·

Incubated for 10 minutes at ambient temperature. The reaction was initiated by the addition of 5 µl of [ ¹⁴ C] arachidonic acid from the ethanol pool (final concentration = 9.5 mM) and then proceeded for 3 minutes. The reaction was quenched by the addition of 0.2 mL of ethyl ether: methanol: 0.2 M citric acid (30: 4: 1) followed by centrifugation of the organic phase at 10,000 µg for 1 minute. 50 mL of the organic phase was transferred with a glass capillary pipette to silica gel 60A TLC plates (Whatman # LK6D). The plates were developed in petroleum ether: ethyl ether: acetic acid (15: 85: 0.1) at ordinary temperature for 30 minutes. Plates were exposed to Kodak XAR-5 film for 24 hours. The film was developed, measured with a densitometer, and peak areas belonging to arachidonic acid and its products were calculated. Percent inhibition was determined from the amount of arachidonic acid converted to oxidation products in samples containing test substance relative to control samples (no test substance).

Table 5 provides data on inhibition of soluble 5-lipoxygenase in RBL-1 cell extract by racemic compound 202 as well as its enantiomers and compounds 216,217, 234 and 236.

Example 9 Inhibition of leukotriene B4 production in ionophore-stimulated human blood

Human blood was transferred to heparinized blood collection tubes and then transferred in 1 ml aliquots into microfuge tubes. 5 ml of the test compound at various concentrations were dissolved in DMSO and the solution was then added to the blood sample and allowed to incubate for 30 minutes at 37 ° C. Then calcium ionophore (5 ml) (A23187) in DMSO was added to a final concentration of 50 mM and the samples were incubated for 30 minutes at 37 ° C. The samples were then centrifuged at 1,100 µg (2500 rpm, rotor H1000B, Sorvall centrifuge) for 10 minutes at 4 ° C. 100 ml of the supernatant was transferred to a 1.5 ml microfuge tube and 400 ml of cold methanol was added and the protons precipitated on ice for 30 minutes. The samples were further centrifuged at 110 x g for 10 minutes at 4 ° C and the supernatant was assayed for LTB4 using a commercially available EIA kit (Cayman Chemical) according to the manufacturer's instructions.

Ti · t · tt · «« «

I · v ** 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

Table 5 provides data on the inhibition of leukotriene B4 production in ionophore-stimulated human blood by racemic compound 202 as well as its enantiomers, as well as by compounds 216, 217,234 and 236.

Example 9 Evaluation of 5-lipoxygenase ex-vivo in mice and blood lipoxygenase in rats

Female CD-I mice weighing 18-25 g and female CD rats weighing 150-230 g were obtained from Carles River Labs. Compounds were dissolved in 0.5% DMSO in 0.9% NaCl for administration to mice (0.5 mg / L) and in an alcoholic carrier (2% benzyl alcohol, 1% ethanol, 40% PEG 300.10% propylene glycol, 47%). % 5% dextrose plus 3.5% pluroniv F-68 in DI20) for administration to rats. The animals were injected with compound (0.5% DMSO in saline, 10 ml / kg in mice; alcohol carrier, 1 ml / kg in rats) minutes before decapitation. Heparinized blood (0.3 ml) was added to 1.5 ml Eppendorf centrifuge tubes containing 3 ml of 2 mM calcium ionophore A23187 (final concentration of A23187 was 20 mM). The sample was incubated for 30 minutes in a 37 ° C water bath and then centrifuged for 2 minutes. Plasma was diluted (χ 120) and examined for LTB ₄ using ElA.

Table 5 provides data on ex-vivo 5-lipoxygenase values in mouse blood lipoxygenase in rats after administration of racemic compound 202 as well as its enantiomers, and compounds 216, 217, 234 and 236.

Example 10 Glucuronidation ratio

The glucuronidation ratio is a measure of the metabolic stability in vivo of the compounds disclosed herein.

Glucuronidation reactions were performed with a reaction mixture containing 2 mg / ml human microsomal protein, 5 mM magnesium chloride 100100 mM Tris HCl (pH 7.4), 0.1-1.0 mM substrate, and 3 mM UDP-glucuronic acid. After incubation at 37 ° C for 0 (control), 15,30,45,60,90,120, 180 and 240 minutes, 40 μΐ aliquots of the reaction mixture were mixed with 80 μΐ acetonitrile and centrifuged to remove precipitated proteins. Aliquots of the supernatant were analyzed by HPLC

44 ·· • 4 · 4 · ·

4 4 · 4 • «43 4

4 4 4

445 44 44 «·· • 4 44

4 4 4

4 44

443 4 4

4 4

44 with reverse phase to determine parent compound death and metabolite formation.

Table 5 provides data, Figure 2 illustrating, the glucuronidation ratio of racemic compound 202 as well as its enantiomers, and compounds 216,217, 234 and 236.

Figure 3 illustrates the glucuronidation ratio for the displayed enantiomers.

Example 11 Effect of Compounds on Cytosolic 5-Lipoxygenase of Rat Basophil Leukemia Cells

RBL-2H3 cells were grown to confluence in tissue culture flasks, according to Carter et al. (J. Pharm. Exp Ther 256 (3); 929-937, 1991). Cells were harvested and washed five times in D-PBS without calcium and magnesium. Cells were suspended at 2 x 10 ⁷ / ml in 10 mM BES, 10 mM BES, 10 mM PIPES, pH 6.8.1 mM EDTA, and then sonicated. The sonicate was centrifuged at 20,000 µg for 20 minutes at 4 ° C. The supernatant was removed and stored in aliquots at -70 ° C.

The 5-LO activity in RBL-2H3 was determined as follows: 0.1 ml reaction mixture consisting of 0.7 mM CaCl ₂ , 100 mM nacL, 1 mM EDTA, 10 mM BES, 10 MM PIPES, pH 7.4, different the concentration of test compound dissolved in DMSO (7.5% DMSO as the final concentration in the experiment) and from that amount of RBL-2H3 that converted 15% of the arachidonic acid substrate mixture to oxidized products (determined experimentally for each RBL-2H3 preparation) were incubated for 20 minutes at normal temperature. The reaction was initiated by adding 5 µl of arachidonic acid substrate mixture (0.994 nmol [ ¹⁴ C] arachidonic acid and 6.06 nmol arachidonic acid per experiment in NH 4 OH) and allowed to run for 5 minutes at 37 ° C. The reaction was terminated by the addition of 0.12 mL of (i) 1.66 mg / mL triphenylphosphine in ethyl ether; (ii) methanol; and (iii) 0.2 M citric acid (30: 4; 1), followed by centrifugation at 1000 µg for 1 minute. 50 µl of the organic phase was transferred by glass capillary to TLC plates 60A (Whatman # 6KDF). Plates were developed in ethyl ether: acetic acid (100: 0.1) for 25 minutes at ambient temperature. Plates were exposed to Kodak X-OMAT film for 40 hours. The film was developed, measured with a densitometer, and the areas of the acid-related peaks were counted. 4 4 ♦ ··

4 4 4 4 4

44443 4 4

4 4 4 4 «4« 44 44 444

4 4

4 4

4 4 • 4 4

44. 04 arachidone and its products. Percent inhibition was calculated from the amount of [ ¹⁴ C] arachidonic acid converted to oxidized products in samples of test compound relative to control (no test compound).

The results are shown in Table 4.

Example 12 Inhibition of leukotriene B4 production in ionophore-stimulated human blood Human blood was transferred to heparinized blood collection tubes and then transferred in 1 ml aliquots into micro-fuged tubes. 5 ml of the test compound at various concentrations were dissolved in DMSO and the solution was then added to the blood sample and allowed to incubate for 30 minutes at 37 ° C. Then calcium ionophore (5 ml) (A23187) in DMSO was added to a final concentration of 50 mM and the samples were incubated for 30 minutes at 37 ° C. The samples were then centrifuged at 1100 x g (2500 rpm, rotor.10000, Sorvall centrifuge) for 10 minutes at 4 ° C. 100 ml of the supernatant was transferred to a 1.5 ml microfuge tube and 400 ml of cold methanol was added and the protons were precipitated on ice for 30 minutes. The samples were further centrifuged at 110 x g for 10 minutes at 4 ° C and the supernatant was assayed for LTB4 using a commercially available EIA kit (Cayman Chemical) according to the manufacturer's instructions.

The results are shown in Table 5.

a * * in »

aaa aa aa aa aa aa aa aa

TABLE 4

RBL 1 HWB I esc vivo L. TB4

· · Ft ft ft • ft ft ft * ft ft · i ft

ft ft ft ft ft ft ft ft ft ft ft ft ft ft ft ft

TABLE 4 (continued)

Compound 406 a

fe fe ··· · a · k · · i ··· ··

Example 4 Evaluation of ex-vivo mouse blood 5-lipoxygenase

Female CD-I mice weighing 18-25 g and female CD rats weighing 150-230 g were obtained from Carles River Labs. Compounds were dissolved in 0.5% DMSO in 0.9% NaCl for administration to mice (0.5 mg / L) and alcohol carrier (2% benzyl alcohol, 1% ethanol, 40% PEG 300.10% propylene glycol, 47%). % 5% dextrose plus 3.5% pluroniv F-68 in DiH ₂ O) for administration to rats. The animals were injected with compound (0.5% DMSO in saline, 10 ml / kg in mice; alcohol carrier, 1 ml / kg in rats) minutes before decapitation. Heparinized blood (0.3 ml) was added to 1.5 ml Eppendorf centrifuge tubes containing 3 ml of 2 mM calcium ionophore A23187 (final concentration of A23187 was 20 mM). The sample was incubated for 30 minutes in a 37 ° C water bath and then centrifuged for 2 minutes. Plasma was diluted (x120) and screened for LTB4 using EIA.

The results are shown in Table 5.

Example 5 Glucuronidation studies

The glucuronidation ratio is a measure of the metabolic stability in vivo of the compounds disclosed herein.

Glucuronidation reactions were performed with a reaction mixture containing 2 mg / ml human microsomal protein, 5 mM magnesium chloride, 100 mM Tris HCl (pH 7.4), 0.1-1.0 mM substrate and 3 mM UDP-glucuronic acid. After incubation at 37 ° C for 0 (control), 15, 30,45, 60,90, 120,180 and 240 minutes, 40 μΐ aliquots of the reaction mixture were mixed with 80 μΐ acetonitrile and centrifuged to remove precipitated proteins. Aliquots of the supernatant were analyzed by reverse phase HPLC to determine parent compound death and metabolite formation.

The results are shown in Figure 5.

Example 6 Evaluation of eosinophilic infiltration

The accumulation of inflammatory cells in the lungs is one of the pathological phenomena associated with asthma. Increase in leukocyte levels, especially eosinophils in blood and lung fl 4 • 4 • · 4

4Í · *

fc «3»

• 4 4 i »4

4 ·

4b * «4

4 4 · • 4 4 4 4

4 4 4 4

Fluid was observed after allergen inhalation. Eosinophils have been shown to be important effector cells in allergic inflammations and have cytotoxic properties with respect to their granule proteins and the potential to release inflammatory mediators. Prevention of allergen-induced eosinophils is considered an important point in the development of new antiasthmatic drugs.

Leukotrienes are products of arachidonic acid metabolism by 5-lipoxygenase (5-LO). Lipoxygenase metabolites (LTB ₄ , 5-oxo-15-hydroxyicosatetraenoic acid) have been identified to have a strong ability to recruit eosinophils. A 5-L0 inhibitor that is capable of blocking the immediate brontichide as well as the later accumulation of eosinophils in lung tissue may be beneficial for the prevention and treatment of asthma.

Infiltration of eosinophils into the lungs can be measured by counting the number of cells in bronchoalveolar fluid (BALF) from guinea pigs and mice exposed to allergens.

Guinea pig model. Female Hartley Guinea pigs weighing 400-500 g were actively sensitized to ovalalbumin (OVA) ip by injecting 20 µg OVA and 100 mg Al (OH) ₃ in 0.5 ml 0.9% NaCl on day 1 and day 2. Animals was exposed to 0.5% OVA (in 0.9% NaCl) for 30 seconds on day 15 and day 16. Compounds were prepared in 10% PEG or 0.5% carboxymethylcellulose and administered three times (1 hour before exposure). influence and between two such influences). To prevent death due to histamine release, pyrilamine (3 mg / kg, ip) was administered 15 minutes prior to exposure. After 24 hours following the first exposure (or 4 hours after the last exposure), the animals were allowed to bleed under a neck anesthetic. BAL was performed with 2 x 10 ml 0.5 mM EDTA in DPBS (w / o Ca ²⁺ , Mg ²⁺ ) at 37 ° C by tracheal cannulation. The total number of cells in BAL fluid was measured with a Sysmex microcell counter (F-800) and the differential cells were counted on a cytospin preparation. Percent inhibition on total cell count or eosinophil accumulation = [(blank) - (blank)] / (blank) x 100.

Mouse model: Male C57 BL / 6 mice, weighing 21-23 g, were actively sennised to oVA by administering 10 µg OVA and 1 mg A1 (OH) ₃ in 0.2 ml 0.9% NaCl on day 1. Hypersensitivity developed following a daily inhalation aerosol of 1% OVA or saline for 30 minutes between days 14 and 21.

«*

«· * ··

β · «φ

V φ

♦ · · ·

Compounds were prepared in 10% PEG 200 or 0.5% carboxymethylcellulose and administered orally at doses of 20 mg / kg, bid between days 18 and 22. Animals were allowed to bleed under neck anesthetics four hours after the last inhalation of OVA. BAL was performed with 2 x 10 ml 0.5 mM EDTA in DPBS (w / o Ca ²⁺ , Mg ²⁺ ) at 37 ° C by tracheal cannulation. Total cell counts in BAL fluid were measured with a Sysmex microcell counter (F800) and differential cells were counted on a cytospin preparation. Percent inhibition on total cell count or eosinophil accumulation = [(blank) - (blank)] / (blank) x 100. (See Yeadon M. et al. Agents Actions 38: 8-17, 1993; Brusselle GG et al. ALA'94, A754; ScwenkU et al. J. Biol. Biochem. 267: 12482-12488, 1992; and Clinic M et al. Cur. Opin. Immunol. 6; 860-864, 1994 ).

.. .t, h. ..- rfv.T-whlMM · ib

Table 5

STRUCTURE

ÓQ1 «uU

RBL

Xinh.

IC 50 <»· nU µM

HWB% inh. icsa · ft ft a a a a a a a a a a

S · · · · · ft ft ft ··· "4 * ft ft ft · ·« ^{ft 4} »·« · «ftft * in ·« · <3o »« mftk the ex vivo LTB4% Wv tcso

207 1.4 2fl 2000 0.4 · RAT : Μ® 04.4V Mon 04, (40 * Mo 43.40 M «34340 pXy ^^^ Ar ' ¹ ' « ^Ϊ Α> SSR T> 93 * 4« C ”» ⁰ 220 moo 0.74 RAT M ««, «* Mo 57. (40 * M> 343 TK.d ^ CH ₃ ⁰ 223 ^ 2W4e ^J 3SM 032 AND RAT Mo 40.40 * M 52.140 * M 44. (40 * 24 »12J40 *

STRUCTURE

RBL

HWB exvivo LTB4% W *. K ^ Q

Oo »moflt • a a« Μ

Table 5

4 «* · (44

ICSO to ftW wM

, 234'Ϋ'ΧΓ _t 234 ”*** 3100 3Λ RAT Xpo 73.00 24 »« 3,110 24 »± 0.309 Xr2 ^ Tf · ' 236 2000 0.73 RAT 2.po 42,09 2jx> -4,109 24 »« A «e 24 »-1,109 010 SM RAT J> * 07.00 * 3> 22.129 »4 »« Μ 104 »03-109 '

»♦

ti 9 • 9 · ♦♦ ·

* ♦ · • • • • • • • • •

STRUCTURE

RBL

Ex vivo HWB LTB4 d4M µM% W ».

IC50ddM nM uM% Wi IC50 to ·· ^{% lnh} · KSO

0 1 too 0.007 RAT 2, pq 61.60 * 2, M 7.2.140 2. »63.W 2j »37.1» 0 0.143 Xn9 ~ - & 4.145 0 XrQ · —ť 0.17 »

««

Φ

Φ

ΦΦΦ · * * * * * * * *

ΦΦ · · · Φ

Modifications and variations of the present invention relating to compounds that reduce the formation of oxygen radicals during respiratory or immune responses will be apparent to those skilled in the art. Such modifications and variations are set forth in the appended claims.

Claims (5)

(Claim! 1. Compounds of formula: (Y) (I) where: Ar is an aryl or heteroaryl group, which is optionally substituted with at least one group selected from the group consisting of halogen (including but not limited to fluorine), lower alkoxy (including methoxy), lower aryloxy (including phenoxy), W, cyano or R; m is 0; W is independently -AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , AN (OM) C (O) R ⁴ , -AC (O) N (OM) R ^4, -C (O) N (OM) R ^4, -C (O) NHA or -AB; A is lower alkynyl, alkaryl or aralkyl wherein one or more carbons may be replaced by O, N or S; B is selected from the group consisting of pyridylimidazole and benzimidazole, each of which is optionally substituted with R ³ ; M is hydrogen, a pharmaceutically acceptable cation or metabolically cleavable group; X is O, S, S (O), NR ⁵ or CHR ⁵ ; Y is O, S, S (O), NR ⁵ or CHR ⁵ ; R ¹ and R ² are "independent hydrogen, lower alkyl including methyl-cyclopropyimethyl", ethyl, isopropyl, butyl, pentyl, hexyl and C 3-8 cycloalkyl such as cyclopentyl; halogen lower alkyl such as trifluoromethyl; halogen and -COOH; R ³ and R ⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl, aralkyl, alkaryl, C 1-6 alkoxy-C 1-10 alkyl, C 1-6 alkylthio-C 1-10 alkyl, heteroaryl or heteroarylalkyl; 4 * fl 4 »* 4 •• 444 in 4 4 4 4 · «« · · 4 4 4 4 4 4 4 43 «« 44 * ♦ 4 · 4 * ·· R ⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, alkarylAN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , - AN (OM) C (O) R ⁴ , -AC (O) N (OM) R ⁴ , AS (O) n R ³ , -AS (O) n CH 2 C (O) R ³ , -AS (O) n CH 2 CH ( OH) R ³ , -AC (O) NHR ³ and wherein n is 0 -2. The compound of claim 1, wherein Ar is selected from the group consisting of phenyl, trimethoxyphenyl, dimethoxyphenyl, fluorophenyl and in particular 4-fluorophenyl, difluorophenyl, pyridyl, dimethoxypyridyl, quinolinyl, furyl, imidazolyl and thienyl. The compound of claim 1, wherein -AB is and wherein Ar is aryl or heteroaryl substituted with at least one group selected from W, halogen, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acids, sulfate, acids phosphonate, phosphate and phosphonate. A pharmaceutical composition comprising an effective amount of the claimed compound 1, 2, 3, 4, 5, 9 or 10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. A method for treating inflammatory conditions in a patient receiving an effective amount of Compound 1, 2, 3, 4, 5, 9, 10 or a pharmaceutically acceptable salt thereof. The method of claim 7, wherein the animal is selected from human, savx, horse, dog, and bovine. 9. Compound of formula: R ¹ R ² Ar (I) X fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe fe. fefe fefe ¹ ·· fefe fefe fe fefe fefe · fefe fe where: Ar is an aryl or heteroaryl group, which is optionally substituted with at least one group selected from the group consisting of halogen (including but not limited to fluorine), lower alkoxy (including methoxy), lower aryloxy (including phenoxy), W, cyano or R ³ ; m is 0; W is independently -AN (R ³ ) C (O) N (OM) R ⁴ , -AN (OM) C (O) R ⁴ , -AC (O) N (OM) R ⁴ , C (O) N ( OM) R ⁴ , -C (O) NHA or -AB; A is lower alkyl, lower alkenyl, lower alkynyl, alkaryl or aralkyl, wherein one or more carbons may be replaced by O, N or S; B is selected from the group consisting of pyridylimidazole and benzimidazole, each of which is optionally substituted with R ³ ; M is hydrogen, a pharmaceutically acceptable cation or metabolically cleavable group; X is O, S, S (O), NR ⁵ or CHR ⁵ ; Y is O, S, S (O), NR ⁵ or CHR ⁵ ; R ¹ and R ² are independently hydrogen, lower alkyl including methyl, cyclopropylmethyl, ethyl, isopropyl, butyl, pentyl, hexyl, and C ₃ _ ₈ cycloalkyl, e.g. cyclopentyl; halogen lower alkyl such as trifluoromethyl; halogen and -COOH; R ³ and R ⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl, aralkyl, alkaryl, C 1-6 alkoxy-C 1-10 alkyl, C 1-6 alkyl; ₆ alkylthio - C] .10 alkyl, heteroaryl or heteroarylalkyl; R ⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, alkaryl, AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , - AN (OM) C (O) R ^4, -AC (O) N (OM) R ^4, S (O) n R ^3, -AS (O) NCH2C (O) R ^3, -AS (O) n CH ₂ CH (OH) R ³ , -AC (O) NHR ³ and wherein n is 0 -2. 10. A compound of formula Af (I) • ♦ · · · · · φ φ φ φ ΦΦ φ ΦΦ ΦΦ · · · ΦΦ · '· * · * · * · Φ · Φ · * · * · * · * · kde · kde kde kde kde kde kde kde kde kde kde Ar is an aryl or heteroaryl group, which is optionally substituted with at least one group selected from the group consisting of halogen (including but not limited to fluorine), lower alkoxy (including methoxy), lower aryloxy (including phenoxy), W, cyano or R ³ ; m is 1; W is independently -AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , AN (OM) C (O) R ⁴ , -AC (O) N (OM) R ^4, -C (O) N (OM) R ^4, -C (O) NHA or -AB; A is lower alkyl, lower alkenyl, lower alkynyl, alkaryl or aralkyl, wherein one or more carbons may be replaced by O, N or S; B is selected from the group consisting of pyridylimidazole and benzimidazole, each of which is optionally substituted with R ³ ; M is hydrogen, a pharmaceutically acceptable cation or metabolically cleavable group; X is O, S, S (O) or NR ⁵ ; Y is O, S, S (O), NR ⁵ or CHR ⁵ ; R ¹ and R ² are independently hydrogen, lower alkyl including methyl, cyclopropylmethyl, ethyl, isopropyl, butyl, pentyl, hexyl and C ₃ . ₈ cycloalkyl such as cyclopentyl; halogen lower alkyl such as trifluoromethyl; halogen and -COOH; R ³ and R ⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl, aralkyl, alkaryl, C 1-6 alkoxy-C 1-10 alkyl, C 1 alkylthio-C 1-10 alkyl, heteroaryl or heteroarylalkyl; R ⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, alkaryl, AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , - AN (OM) C (O) R ^4, -AC (O) N (OM) R ^4, S (O) n R ^3, -AS (O) 'CH 2 C (O) R ^3, -AS (O) n CH ₂ CH (OH) R ³ , -AC (O) NHR ³ and wherein n is 0 -2. 11. Compound 6 of formula OOm, w (i) • »♦ '.'4 · 4 4 4 4 5 · »· * 4 4 4 »4 44 4 «* 3 4 4 4 '4 4 4 • 4 444 4 4 4 4 where: Ar is an aryl or heteroaryl group, which is optionally substituted with at least one group selected from the group consisting of halogen (including but not limited to fluorine), lower alkoxy (including methoxy), lower aryloxy (including phenoxy), W, 3 * ^f cyano or R; m is 0 or 1; W is independently -AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , TT T. A AN (OM) C (Q) R 1 = AC (U) N (UM) K 1, -L (U) N (UM) R 6, -C (U) NHA or -A-hi; A is lower alkyl, lower alkenyl, lower alkynyl, alkaryl or aralkyl, wherein one or more carbons may be replaced by O, N or S; B is selected from the group consisting of pyridylimidazole and benzimidazole, each of which is optionally substituted with R ³ ; M is hydrogen, a pharmaceutically acceptable cation or metabolically cleavable group; X is O, S, S (O), NR ⁵ or CHR ⁵ ; Y is O, S, S (O), NR ⁵ or CHR ⁵ ; R and R are independently hydrogen, lower alkyl including methyl, cyclopropylmethyl, ethyl, isopropyl, butyl, pentyl, hexyl and C 3-8 cycloalkyl such as cyclopentyl; halogen lower alkyl such as trifluoromethyl; halogen and -COOH; R ³ and R ⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl, aralkyl, alkaryl, C 1-6 alkoxy-C 1-10 alkyl, C 1-3 alkylthio-C 1-6 alkyl; Alkyl, heteroaryl or heteroarylalkyl; R ⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, alkaryl, AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , - AN (OM) C (O) R ^4, -AC (O) N (OM) R ^4, S (O) n R ^3, -AS (O) NCH2C (O) R ^3, -AS (O) n CH ₂ CH (OH) R ³ , -AC (O) NHR ³ and wherein n is 0 -2. A pharmaceutical composition comprising an effective amount of the claimed Compound 11 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. A method for treating inflammatory conditions in a patient receiving an effective amount of Compound 11 or a pharmaceutically acceptable salt thereof. The compound of claim 1,9 or 10, wherein X is O. The compound of claim 1,9 or 10, wherein the aryl group has a halogen. • φφ · Β « Φ Φ Φ Φ · · · · · · · ♦ Φ Φ * Φ • Φ Φ Φ • Φ « φφ Φ Φ Φ · Φ • Φ ΦΦ • 9 Φ Φ Φ Φ Φ Φ ΦΦ Φ Φ ΦΦ «· The compound of claim 1 or 10, wherein W is AN (OM) C (O) NR ³ R ⁴ . The compound of claim 16, wherein R ³ and R ^{4 are} hydrogen atoms. The compound of claim 1,9 or 10, wherein R and R are hydrogen atoms. The compound of claim 1, wherein A is lower alkynyl. The compound of claim 1, wherein Μ is H. The compound of claim 11, wherein X is O. The compound of claim 11, wherein the aryl group has halogen. Ot ~, Z__1 ___. ». '·. 1, z, j. Línjuccmiiu utuujs.u zz, in Kierc is naiogen nuor. The compound of claim 23, wherein the fluorine is in the para position. The compound of claim 11, wherein W is AN (OM) C (O) NR ³ R ⁴ . The compound of claim 25, wherein R ³ and R ^{4 are} hydrogen atoms. The compound of claim 11, wherein R ¹ and R ^{2 are} hydrogen atoms. The compound of claim 5, wherein A is alkyl. The compound of claim 28 wherein the lower alkyl is butyl The compound of claim 11, wherein the lower alkyl is isopentyl. The compound of claim 1 wherein Μ is H. A compound of the formula wherein? Ar is an aryl or heteroaryl group, which is optionally substituted with at least one group selected from the group consisting of halogen (including but not limited to fluorine), lower alkoxy (including methoxy), lower aryloxy (including phenoxy), W, cyano or R; m is 0 or 1; 99 99 * 9 · 9 9 9 9 9,999 9 * 9 9 9 9 * 99 99 99 9 99 99 98 9 9 9 9 9 9 9 99 9 9 9 9 »99 99 99 'is 1-6; W is independently -AN (OM) C (O) N (R ³ ) R ⁴ , -N (OM) C (O) N (R ³ ) R ⁴ , AN (R ³ ) C (O) N (OM) R ^4, -N (R ³⁾ C (O) N (OM) ^R4, -AN (OM) C (O) R ^4, -N (OM) C (O) R ^4, -AC (O) N (OM) R ^4, -C (O) N (OM) R ^4, -C (O) NHA; A is lower alkyl, lower alkenyl, lower alkynyl, alkylaryl or arylalkyl, wherein one or more carbons may be replaced by O, N or S (with valency supplemented with hydrogen and oxygen if necessary) but -YA, -A- or -AW- should not contain two 8- - —-adjacent heteroatoms (e.g., Θ-Θ -, -S-S-, -G-Ss-aid); ' M is hydrogen, a pharmaceutically acceptable cation or metabolically cleavable group; > X is O, S, S (O), S (O) ₂ , NR ⁵ or CHR ⁵ ; Y is O, S, S (O), S (O) ₂ , NR ^5, or CHR ⁵ ; Z is O, S, S (O), S (O) ₂ , NR ⁵ ; . R ¹ and R ² are independently hydrogen, lower alkyl including methyl, cyclopropylmethyl, ethyl, isopropyl, butyl, pentyl, hexyl, and C ₃ .g cycloalkyl, e.g. cyclopentyl; halogen lower alkyl such as trifluoromethyl; halogen and -COOH; R ³ and R ⁴ are independently hydrogen or alkyl, alkenyl, alkynyl, aryl, arylalkyl, alkylaryl, C 1-6 alkyl; ₆ alkoxy-C io alkyl, Ci_e alkylthio - C -i ₀ alkyl, heteroaryl, or% heteroarylalkyl; R ⁵ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, arylalkyl, alkylaryl, AN (OM) C (O) N (R ³ ) R ⁴ , -AN (R ³ ) C (O) N (OM) R ⁴ , -AN (OM) C (O) R ^4, -AC (O) N (OM) R ^4, S (O) xR ^3, -AS (O) NCH2C (O) R ^3, -AS (O) nCH ₂ CH (OH) R ³ , -AC (O) NHR ³ and wherein x is 0 -2. The compound of claim 32, wherein Ar is selected from the group consisting of phenyl, trimethoxyphenyl, dimethoxyphenyl, fluorophenyl, difluorophenyl, pyridyl, dimethoxypyridyl, quinolinyl, furyl, imidazolyl and thienyl. The compound of claim 32, wherein Ar is 4-fluorophenyl. The compound of claim 32, wherein Z is O. The compound of claim 32, wherein Z is S. The compound of claim 32, wherein - (Y) _m W is selected from the group consisting of: • ·· '·· · ·· ·· «51 IN The compound of claim 2, wherein - (Y) _m W is selected from the group consisting of: NHj The compound of claim 32, 33, 34,35,36,37 or 38 in at least 97% enantiomerically enriched form. ~. 40. A compound selected from the group consisting of 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4-protected oxy-1-butynyl) tetrahydrofuran and 2S, 5R-cis-2- (4-fluorophenoxymethyl) -5- ( 4-protected oxy-1-butynyl-tetrahydrofuran. The compound of claim 32 which is 2S, 5R-trans-2- (4-fluorophenoxymethyl) -5- (4N-hydroxyureidyl-1-butynyl) tetrahydro furan. • i * «·::::: * * * ** ** ** ** ** ** ** ** ** ** ** ** · «« ** ** ** ** ** ** ** The compound of claim 32 which is 2S, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4N-hydroxyureidyl-1-butynyl) tetrahydrofuran. The compound of claim 32 which is 2R, 5S-trans-2- (4-fluorophenoxymethyl) -5- (4N-hydroxyureidyl-1-butynyl) tetrahydrofuran. The compound of claim 32 which is 2R, 5R-trans-2- (4-fluorophenoxymethyl) -5- (4N-hydroxyureidyl-1-butynyl) tetrahydro furan. A pharmaceutical composition comprising an effective amount of a compound of claim 32 -44, or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable carrier. Any compound of the above claims for use in medical therapy, eg, for the treatment of prophylaxis of inflammatory disorders. 47. The use of any of the above requirements and pharmaceutically acceptable derivatives or salts in the manufacture of a medicament for the treatment of inflammatory disorders. 48. Any compound claimed above in combination with a pharmaceutically acceptable carrier.